Pyridine derivatives

ABSTRACT

Pyridine compounds of general formula:wherein -R1 representsin which R11 is hydrogen, C1-6 alkyl, halogen, hydroxy, C1-12 alkoxy, nitro, amino, C1-6 alkylsulfonylamino, C1-6 alkoxycarbonyl, C1-6 alkylamino, di(C1-6 alkyl)amino, C1-6 alkanoylamino, phenyl C1-6 alkylamino, phenylsulfonylamino, or -O-(CH2)n-R111; R2 represents hydrogen or halogen; R3 represents hydrogen, -CR31R32R33, or -NR34R35; R4 is hydrogen, carbamoyl, CN, carboxyl, etc.; R5 is amino, C1-6 alkylamino, di C1-6 alkylamino, etc. or salt thereof. The compound has an excellent anti-inflammatory activity, and other biological activity.

TECHNICAL FIELD

The present invention relates to novel pyridine derivatives, processesfor preparing them and pharmaceutical preparations containing them. Thepyridine derivatives of the present invention inhibit IκB kinase β(IKK-β or IKK-beta) activity, thus inhibit nuclear factor kappa B(NF-κB) activity, and can be used for the prophylaxis and treatment ofdiseases associated with NF-κB activity, in particular for the treatmentof inflammatory diseases.

BACKGROUND ART

Nuclear factor kappa B (NF-κB) belongs to a family of closely relatedhomo- and hetero-dimeric transcription factor complexes composed ofvarious combinations of the Rel/NF-κB family of polypeptides. NF-κB andrelated family members are involved in the regulation of more than 50genes relating to immune and inflammatory responses ((Barnes P J, KarinM (1997) N Engl J Med 336, 1066-1071) and (Baeuerle P A, Baichwal V R(1997) Adv Immunol 65, 111-137)). In most cell types, NF-κB is presentas a heterodimer comprising a 50 kDa and a 65 kDa subunit (p50/RelA).The heterodimer is sequestered in the cytoplasm in association withinhibitor of NF-κB (IκB)-family of proteins to be kept in an inactivestate. IκB-family proteins mask the nuclear translocation signal ofNF-κB. Upon stimulation of cells with various cytokines (e.g. TNF-α,IL-1), CD40 ligand, lipopolysaccharide (LPS), oxidants, mitogens (e.g.phorbol ester), viruses or many others. IκB proteins are phosphorylatedat specific serine residues, poly-ubiquitinated, and then degradedthrough a proteasome-dependent pathway. Freed from IκB, the active NF-κBis able to translocate to the nucleus where it binds in a selectivemanner to preferred gene-specific enhancer sequences. Among the genesbeing regulated by NF-κB are many coding for pro-inflammatory mediators,cytokines, cell adhesion molecules, and acute phase proteins. Expressionof several of these cytokines and mediators in turn can lead to furtheractivation of NF-κB via autocrine and paracrine mechanisms.

Broad evidence is available that suggests a central role of NF-κB inmany inflammatory disorders including airway inflammation and asthma((Yang L et al., J Exp Med 188 (1998), 1739-1750), (Hart L A et al. Am JRespir Crit Care Med 158 (1998), 1585-1592), (Stacey M A et al., BiochemBiophys Res Commun 236 (1997), 522-526) (Barnes P and Adcock I M, TrendsPharmacol Sci 18 (1997), 46-50)).

Further, it has been shown that glucocorticoids, which are by far themost effective treatment for asthma, inhibit airway inflammation bydirectly interacting with and inhibiting the activity of thetranscription factors NF-κB and activating peptide-1 (AP-1) ((Barnes P(1997) Pulmon Pharmacol Therapeut 10, 3-19) and (Dumont A et al. (1998)Trends Biochem Sci 23, 233-235)).

In general, inhibition of NF-κB activation results in stronganti-inflammatory effects similar or superior to those brought upon bysteroids. Consequently, NF-κB inhibition should improve inflammatorysymptoms typical for asthma; allergic rhinitis; atopic dermatitis;hives; conjunctivitis; vernal catarrh; rheumatoid arthritis; systemiclupus erythematosus; psoriasis; diabrotic colitis; systemic inflammatoryresponse syndrome; sepsis; polymyositis; dermatomyositis; Polyaritisnodoa; mixed connective tissue disease; Sjoegren's syndrome; gout, andthe like.

Further, several studies imply that NF-κB plays an essential role inneoplastic transformation. For example, NF-κB is associated with celltransformation in vitro and in vivo as a result of gene overexpression,amplification, rearrangement, or translocation (Mercurio, F., andManning, A. M. (1999) Oncogene, 18:6163-6171). In certain human lymphoidtumor cells, the genes of NF-κB family members are rearranged oramplified. Its possible involvement in cancer pathology is alsodisclosed in Mayo, M. W., Baldwin A. S. (2000) Biochmica et BiophysicaActa 1470 M55-M62. Mayo M. W. et al., discloses the inhibition of NF-κBresults in the blockage the initiation and/or progression of certaincancer, particularly colorectal cancer.

Finally, NF-κB may also be involved in the regulation of neuronal celldeath. It has been shown that NF-κB becomes activated and promotes celldeath in focal cerebral ischemia Nature medicine Vol. 5 No. 5, May1999).

Extensive research during the past years led to the identification of anIκB kinase (IKK) complex as being responsible for the signal-induced IκBphosphorylation ((Mercurio, F., and Manning, A. M. (1999) CurrentOpinion in Cell Biology, 11:226-232), (Mercurio, F., and Manning, A. M.(1999) Oncogene, 18:6163-6171), (Barnkett, M., and Gilmore T. D. (1999)Oncogene 18, 6910-6924), (Zandi, E., and Karin, M., (1999)19:4547-4551), (Israel, A., (2000) trends in CELL BIOLOGY 10:129-133),and (Hatada, E. N, et al. (2000) Current Opinion in Immunology,12:52-58)). This complex is most likely the site of integration of allof the different stimuli leading to NF-κB activation. The IKK-complex(molecular weight 700-900 kDa) is composed of various proteins includingtwo homologous IκB kinases, called IKK-α and IKK-β, an upstream kinase,NIK which induces NF-κB, a scaffold protein called IKAP, which tetherstogether the three kinases, and a regulatory subunit IKK-γ, whichpreferentially interacts with IKK-β.

IKK-β is a 756 amino acid serine-threonine kinase showing 52% identityto and the same domain structure as IKKα ((Mercurio F et al (1997)Science 278, 860-866.), (Woronicz J D et al. (1997) Science 278,866-869.), (Zandi E et al. (1997) Cell 91, 243-252.). IKK-β formshomo-dimers and hetero-dimers with IKK-α in vitro and in cells,respectively. IKK-β also interacts with IKK-γ, IKAP, NIK and IκBα.Recombinant IKK-β phosphorylates IκBα and IκBβ at specific serineresidues with equal efficacy (Li J et al. (1998) J Biol Chem 273,30736-30741.), (Zandi E, Chen Y, Karin M (1998) Science 281,1360-1363.). IKK-β shows a higher constitutive kinase activity ascompared to IKK-α. This is in agreement with data suggesting thatover-expression of IKK-β activates the transcription of aNF-κB-dependent reporter gene with a higher efficacy as compared toIKK-α. IKK-β has been shown to be activated in various cell lines orfresh human cells in response to various stimuli including TNF-α, IL-β,LPS, anti-CD3/anti-CD28 co-stimulation, protein kinase C andcalcineurin, B-cell receptor/CD40 ligand stimulation, and vanadate.IKK-β is activated in fibroblast-like synoviocytes (FLS) isolated fromthe synovium of patients suffering from rheumatoid arthritis orosteoarthritis (Zandi E et al. (1997) Cell 91, 243-252.), (O'Connell M Aet al. (1998) J Biol Chem 273, 30410-30414.), Kempiak S J et al. (1999)J Immunol 162, 3176-3187.). Furthermore, IKK-β can be activated by thestructurally related upstream kinases MEKK-1 and NIK, most likelythrough phosphorylation of specific serine residues within the T-loop(activation loop) and by certain protein kinase C isoforms ((Nakano H etal. (1998) Proc Natl Acad Sci USA 95, 3537-3542.), (Lee F S et al.(1998) Proc Natl Acad Sci USA 95, 9319-9324.), (Nemoto S et al (1998)Mol Cell Biol 18, 7336-7343.), (Lallena M J et al. (1999) Mol Cell Biol19, 2180-2188.)). A catalytically inactive mutant of IKK-β has beenshown to inhibit activation of NF-κB by TNF-α, IL-1β, LPS,anti-CD3/anti-CD28 stimulation ((Mercurio F et al. (1997) Science 278,860-866.), (Woronicz J D et al. (1997) Science 278, 866-869.)). The sameeffects are observed when MEKK1 or NIK are overexpressed. Additionally,IKK-β mutations in the activation loop inhibited IL-1 and TNF-αsignaling (Delhase M et al. (1999) Science 284, 309-313.). Based on theexperimental results described above, there is clear-cut evidence for apivotal involvement of IKK-β in various pathways leading to NF-κBactivation.

In summary, the specific inhibition of IKK-β should result in a stronganti-inflammatory and immuno-modulatory effect in vivo with thepotential of improving the underlying causes of asthma and otherdiseases. In addition, anti-tumor and anti-ischemic effects of an IKK-βinhibitor may be expected.

Manna et al., disclose 4,6-disubstituted 3-cyano-2-aminopyridinesrepresented by general formulas:

wherein (R′, R″) represent (OCH₃, OCH₃), (Cl, Cl), (H, Cl), (H, Br), (H,CH₃), (H, OCH₃), (H, NO₂), or (H, N(CH₃)₂), or

as a general anti-inflammatory, analgesic, and antipyretic agent (Eur J.Med. Chem. 34, 245-254(1999)).

Manna et al. neither disclose pyridine derivatives with aliphatic groupsat position 4 of the pyridine ring, nor suggest IKK-β kinase or NF-κBinhibitory activity on the above known pyridine derivatives.

The development of a novel compound having effective anti-inflammatoryactions based on a specific and selective inhibitory activity to NF-κBhas been desired.

SUMMARY OF THE INVENTION

As the result of extensive studies on chemical modification of pyridinederivatives, the present inventors have found that the compound of novelchemical structure related to the present invention have unexpectedlyexcellent IKK-β kinase inhibitory activity. This invention is to providethe following general formula (I) and the salts thereof:

wherein —R¹ represents

in which —R¹¹ is hydrogen, C₁₋₆ alkyl halogen, hydroxy, C₁₋₁₂ alkoxy,nitro, amino, C₁₋₆ alkylsulfonylamino, C₁₋₆ alkoxycarbonyl, C₁₋₆alkylamino, di(C₁₋₆ alkyl)amino, C₁₋₆ alkanoylamino, phenyl C₁₋₆alkylamino, phenylsulfonylamino, or —O—(CH₂)_(n)—R¹¹¹,

wherein n represents an integer selected from 0 to 6, and R¹¹¹ is C₂₋₆alkenyl, benzoyl, diphenylmethyl, di(C₁₋₆ alkyl)amino, C₁₋₆ alkanoyl,C₁₋₆ alkoxycarbonyl, or a 3 to 10 membered saturated or unsaturated ringhaving 0 to 3 heteroatoms selected from the group consisting of S, O andN as heteroatoms and is optionally substituted by C₁₋₆ alkyl, mono or dihalogen, halogen substituted C₁₋₆ alkyl, nitro, ciano, C₁₋₆alkoxycarbonyl, phenyl, hydroxy, amino, C₁₋₆ alkylamino, di(C₁₋₆alkyl)amino, C₁₋₆ alkanoylamino, C₁₋₆ alkoxy, or carbamoyl;

R² represents hydrogen or halogen;

R³ represents hydrogen or 1,2,3,6-Tetrahydro-pyridine,

—CR³¹R³²R³³,

 wherein R³¹ is hydrogen or C₁₋₆ alkyl,

R³² is hydrogen, α-aminobenzyl, C₁₋₆ alkyl optionally substituted by oneor two substituents selected from the group consisting of hydroxy,amino, amino substituted phenyl, phenyl, halogen substituted phenyl, andC₁₋₆ alkoxy substituted phenyl, or a 5 to 8 membered saturated ringhaving 0 to 3 atoms selected from the group consisting of S, O and N asheteroatoms and optionally substituted by C₁₋₆ alkyl, and

R³³ is hydrogen, amino, C₁₋₆ alkoxycarbonylamino, C₂₋₆alkenyloxycarbonylamino, piperidino-C₁₋₆ alkylcarbonylamino,piperidinyl-C₁₋₆ alkylcarbonylamino, or

R³² and R³³ may form, together with the adjacent carbon atom, a 5 to 8membered saturated ring having 0 to 3 heteroatoms selected from thegroup consisting of N, O and S as heteroatoms, which ring is optionallysubstituted by phenyl-C₁₋₆ alkyl, C₁₋₆ alkoxy substituted phenyl-C₁₋₆alkyl, C₁₋₆ alkyl, amino, ciano, hydroxy, carbamoyl, carboxy, C₁₋₆alkylamino, C₁₋₆ alkoxycarbonyl, di(C₁₋₆ alkyl)amino, benzylamino, C₁₋₆alkylsulfonyl, piperidino C₁₋₆ alkyl carbonyl, or optionally fused bybenzene; or

—NR³⁴R³⁵,

 wherein R³⁴ is hydrogen or C₁₋₆ alkyl and

R³⁵ is hydrogen, a 5 to 8 membered saturated ring having 0 to 3heteroatoms selected from the group consisting of N, O and S asheteroatoms, or —(CH₂)_(m)—NR³⁵¹R³⁵² (m represents any of integers from1 to 6)

wherein R³⁵¹ represents hydrogen, C₁₋₆ alkyl,

R³⁵² represents hydrogen, C₁₋₆ alkyl, C₁₋₆ alkanoyl, C₁₋₆ alkylsubstituted phenyl, benzoyl, C₁₋₆ alkanoyl, phenylaminocarbonyl,phenylsulfonyl, or

R³⁴ and R³⁵ may form, together with the adjacent N atom, a 5 to 8membered saturated heterocyclic ring, and said ring may optionallycontain NH, S or O atom other than the adjacent N atom and optionallysubstituted by carbamoyl, amino, or C₁₋₆ alkyl;

R⁴ represents hydroxycarbonyl, C₁₋₆ alkanoyl, carbamoyl, nitro, cyano,carboxyl, C₁₋₆ alkoxycarbonyl, C₁₋₆ alkylcarbamoyl, C₁₋₆ alkylamino, 5to 10 membered heteroaryl (hydroxy) methyl, 5 to 10 memberedheteroaryl-C₁₋₆ alkyl, or methyl substituted by hydroxy and a 5 to 7membered saturated cyclic ring, C₁₋₆ alkyl optionally substituted by oneselected from the group consisting of hydroxy, C₁₋₆ alkoxy, C₁₋₆alkylsulfonylamino, C₁₋₆ alkylcarbonylamino, C₅₋₁₀ aryl, C₅₋₁₀arylsulfonyl, C₅₋₁₀ arylsulfanyl, C₅₋₁₀ aryloxy, imidazolyl, or dioxosubstituted pyrolidino-oxy,

—(CH₂)_(p)NHCOR⁴¹, —CH₂)_(p)NHC(═S)R⁴¹

 wherein p represents any of integer from 1 to 6 and R⁴¹ represents C₁₋₆alkoxy, amino, phenylamino, C₁₋₆ alkyl, C₁₋₆ alkylamino, di(C₁₋₆alkyl)amino, C₃₋₁₀ cycloalkylamino,

R³ and R⁴ may form, together with the carbon atoms in the pyridine ring,4 to 10 membered monocycloalkyl or bicycloalkyl optionally interruptedby NH and optionally substituted by benzyl, ═NH, or ═O;

R⁵ represents NR⁵¹R⁵²,

wherein R⁵¹ is hydrogen, C₁₋₆ alkyl,

R⁵² is hydrogen, C₁₋₆ alkyl, phenyl, benzyl, C₁₋₆ alkanoyl, or NR51 R52may form saturated 5-6 membered ring optionally contain NH or O as otherheteroatom than the adjacent N atom, or

R⁴ and R⁵ may form,

—R⁴⁰—CO—NH—,

—R⁴⁰—SO₂—NH—,

—R⁴⁰—C(═S)—NH—

—R⁴⁰—CH₂—NH—,

 wherein said —R⁴⁰— represents —CHR⁴⁰¹—O—, —CH₂—NR⁴⁰¹—, —CO—NR⁴⁰¹—,—CH₂—CHR⁴⁰¹—, —CH═CR⁴⁰¹—, (in which R⁴⁰¹ is C₁₋₆ alkanoyl, C₁₋₆ alkyl,phenyl, C₁₋₆ alkylsulfonyl, C₃₋₈ cycloalkylaminocarbonyl, hydrogen,halogen, nitro, amino, ciano, benzoylamino, phenylsulfonyl, carbamoyl,hydroxycarbonyl, C₁₋₆ alkoxycarbonyl, C₁₋₁₂ alkylaminocarbonyl, halogensubstituted C₁₋₆ alkylaminocarbonyl, C₁₋₆ alkanoylamino, C₁₋₆alkylamino, di(C₁₋₆ alkyl)aminocarbonyl, di(C₁₋₆ alkyl)aminoC₁₋₆alkylaminocarbonyl, hydroindenylaminocarbonyl,diphenylmethylaminocarbonyl, pyrrolidinocarbonyl, C₁₋₆ alkoxy C₁₋₆ alkylamino carbonyl, morpholinocarbonyl, piperazinocarbonyl, phenylC₁₋₆alkylaminocarbonyl, hydroxycarbonylC₁₋₆alkyl-aminocarbonyl, C₃₋₈cycloalkylaminocarbonyl, C₃₋₈ cycloalkylC₁₋₆ alkylamino-carbonyl,hydroxyC₁₋₆ alkylaminocarbonyl, carboxyethylaminocarbonyl, C₁₋₆alkylsulfonylaminocarbonyl)

—CR⁴¹═N—NH— (R⁴¹ is hydrogen, amino, or C₁₋₆ alkanoylamino),

—CR⁴²═N—C═N— (R⁴² is hydrogen or amino).

The compounds of the present invention surprisingly show excellent IKK-βkinase inhibitory activity and cytokine inhibitory activity. They are,therefore suitable especially as NF-κB inhibitors and in particular forthe production of medicament or medical composition, which may be usefulto treat NF-κB dependent diseases.

More specifically, since the pyridine derivatives of the presentinvention inhibit IKK-β kinase activity, they are useful for treatmentand prophylaxis of diseases involving NF-κB activity as follows:inflammatory symptoms including asthma; allergic rhinitis; atopicdermatitis; hives; conjunctivitis; vernal catarrh; chronicarthrorheumatism; systemic lupus erythematosus; psoriasis; diabroticcolitis; systemic inflammatory response syndrome (SIRS); sepsis;polymyositis; dermatomyositis (DM); Polyaritis nodoa (PN); mixedconnective tissue disease (MCTD); Sjoegren's syndrome; gout; and thelike.

The compounds of the present invention are also useful for treatment andprophylaxis of diseases like ischemia and tumor, since the diseases alsorelate to IKK-β kinase and NF-κB activity.

Preferred compounds of formula (I) are those wherein:

—R¹ represents

 in which R¹¹ is hydrogen, C₁₋₆ alkyl, halogen, hydroxy, C₁₋₁₂ alkoxy,amino, C₁₋₆ alkanoylamino, phenyl C₁₋₆ alkylamino, phenylsulfonylamino,or —O—(CH₂)_(n)—R¹¹¹,

wherein n represents an integer selected from 1 to 6, and R¹¹¹ is C₂₋₆alkenyl, benzoyl, diphenylmethyl, di(C₁₋₆ alkyl)amino, C₁₋₆ alkanoyl,C₁₋₆ alkoxycarbonyl, or a 3 to 10 membered saturated or unsaturated ringhaving 0 to 3 heteroatoms selected from the group consisting of S, O andN as heteroatoms and is optionally substituted by C₁₋₆ alkyl, mono or dihalogen, halogen substituted C₁₋₆ alkyl, nitro, ciano, C₁₋₆alkoxycarbonyl, phenyl;

R² represents hydrogen;

R³ represents hydrogen, 1,2,3,6-tetrahydro-pyridine

—CR³¹R³²R³³,

 wherein R³¹ is hydrogen or C₁₋₆ alkyl,

R³² is hydrogen, α-aminobenzyl, C₁₋₆ alkyl optionally substituted by oneor two substituents selected from the group consisting of hydroxy,amino, amino substituted phenyl, phenyl, halogen substituted phenyl, andC₁₋₆ alkoxysubstituted phenyl, or a 5 to 8 membered saturated ringhaving 0 to 3 atoms selected from the group consisting of S, O and N asheteroatoms and optionally substituted by C₁₋₆ alkyl, and

R³³ is hydrogen, amino, C₁₋₆ alkoxycarbonylamino, C₂₋₆alkenyloxycarbonylamino, piperidino-C₁₋₆ alkylcarbonylamino, or

R³² and R³³ may form, together with the adjacent carbon atom, a 5 to 8membered saturated ring having 0 to 3 heteroatoms selected from thegroup consisting of N, O and S as heteroatoms, which ring is optionallysubstituted by phenyl-C₁₋₆ alkyl, C₁₋₆ alkoxy substituted phenyl-C₁₋₆alkyl, C₁₋₆ alkyl, amino, carboxy, C₁₋₆ alkylamino, C₁₋₆ alkoxycarbonyl,di(C₁₋₆ alkyl)amino, benzylamino, C₁₋₆ alkylsulfonyl, piperidinoC₁₋₆alkyl carbonyl, or optionally fused by benzene; or

—NR³⁴R³⁵,

 wherein R³⁴ is hydrogen and

R³⁵ is hydrogen, a 5 to 8 membered saturated ring having 0 to 3heteroatoms selected from the group consisting of N, O and S asheteroatoms, or —(CH₂)_(m)—NR³⁵¹R³⁵² (m represents any of integers from1 to 6)

wherein R³⁵¹ represents hydrogen, C₁₋₆ alkyl,

R³⁵² represents hydrogen, C₁₋₆ alkyl, C₁₋₆ alkanoyl, C₁₋₆alkylsubstituted phenyl, benzoyl, C₁₋₆ alkanoyl, phenylaminocarbonyl,phenylsulfonyl, or

R³⁴ and R³⁵ may form, together with the adjacent N atom, a 5 to 8membered saturated heterocyclic ring, and said ring may optionallycontain NH, S or O atom other than the adjacent N atom and optionallysubstituted by carbamoyl, amino, or C₁₋₆ alkyl;

R⁴ represents hydroxycarbonyl, C₁₋₆ alkanoyl, carbamoyl, cyano,carboxyl, C₁₋₆ alkoxycarbonyl, C₁₋₆ alkylcarbamoyl, C₁₋₆ alkylamino, 5to 10 membered heteroaryl (hydroxy) methyl, 5 to 10 memberedheteroaryl-C₁₋₆ alkyl, or methyl substituted by hydroxy and a 5 to 7membered saturated cyclic ring, C₁₋₆ alkyl optionally substituted by oneselected from the group consisting of hydroxy, C₁₋₆ alkoxy, C₁₋₆alkylsulfonylamino, C₁₋₆alkylcarbonylamino, C₅₋₁₀ aryl, C₅₋₁₀arylsulfanyl, C₅₋₁₀ arylsulfenyl, C₅₋₁₀ aryloxy, imidazolyl, or dioxosubstituted pyrolidino-oxy,

—(CH₂)_(p)NHCOR⁴¹, —(CH₂)_(p)NHC(═S)R⁴¹

 wherein p represents any of integer from 1 to 6 and R⁴¹ represents C₁₋₆alkoxy, amino, phenylamino, C₁₋₆ alkyl, C₁₋₆ alkylamino, di(C₁₋₆alkyl)amino, C₃₋₁₀ cycloalkylamino,

R³ and R⁴ may form, together with the carbon atoms in the pyridine ring,4 to 10 membered monocycloalkyl or bicycloalkyl optionally interruptedby NH and optionally substituted by benzyl, ═NH, or ═O;

R⁵ represents NR⁵¹R⁵²,

wherein R⁵¹ is hydrogen, C₁₋₆ alkyl,

R⁵² is hydrogen, C₁₋₆ alkyl, phenyl, benzyl, C₁₋₆ alkanoyl, or NR⁵¹R⁵²may form piperidino, or

R⁴ and R⁵ may form,

—R⁴⁰—CO—NH—, —R⁴⁰—SO₂—NH—,

—R⁴⁰—C(═S)—NH— or

—R⁴⁰—CH₂—NH—,

 wherein said —R⁴⁰— represents —CHR⁴⁰¹—O—, —CH₂-NR⁴⁰¹—, —CO—NR⁴⁰¹—, (inwhich R⁴⁰¹ is hydrogen, C₁₋₆ alkanoyl, C₁₋₆ alkoxycarbonyl, C₁₋₆ alkyl,phenyl, C₁₋₆ alkylsulfonyl, C₃₋₈ cycloalkylaminocarbonyl, C₁₋₆alkylaminocarbonyl, carbamoyl, di (C₁₋₆ alkyl)aminocarbonyl),—CH₂—CHR⁴⁰²—, —CH═CR⁴⁰²—, (in which R⁴⁰² is hydrogen, halogen, nitro,amino, ciano, benzoylamino, phenylsulfonyl, carbamoyl, hydroxycarbonyl,C₁₋₆ alkoxycarbonyl, C₁₋₁₂ alkylaminocarbonyl, halogen substituted C₁₋₆alkylaminocarbonyl, C₁₋₆ alkanoylamino, C₁₋₆ alkylamino, di(C₁₋₆alkyl)aminocarbonyl, di(C₁₋₆ alkyl)aminoC₁₋₆ alkylaminocarbonyl,hydroindenylaminocarbonyl, diphenylmethylaminocarbonyl,pyrrolidinocarbonyl, C₁₋₆ alkoxy C₁₋₆ alkyl amino carbonyl,morpholinocarbonyl, piperazinocarbonyl, phenylC₁₋₆ alkylaminocarbonyl,C₃₋₈ cycloalkylaminocarbonyl, hydroxycarbonylC₁₋₆ alkylaminocarbonyl,C₃₋₈ cycloalkylC₁₋₆ alkylaminocarbonyl, hydroxyC₁₋₆ alkylaminocarbonyl,carboxyethylaminocarbonyl, methylsulfonylaminocarbonyl,)

—CR⁴¹═N—NH— (R⁴¹ is hydroxy, amino, C₁₋₆ alkanoylamino) or

—CR⁴²═N—C═N— (R⁴² is amino)

or a salt thereof.

More preferred compound of formula (I) are those wherein:

—R¹ represents

 in which R¹¹ is hydrogen, C₁₋₁₂ alkoxy, or —O—(CH₂)_(n)—R¹¹¹,

wherein n represents an integer selected from 1 to 6, and R¹¹¹ isphenyl, C₃₋₈ cycloalkyl;

R² represents hydrogen;

R³ represents 1,2,3,6-tetrahydro-pyridine,

—CR³¹R³²R³³,

 wherein R³¹ is hydrogen, and

R³² and R³³ form, together with the adjacent carbon atom, a 5 to 8membered saturated ring interrupted by NH, which ring is optionallysubstituted by phenyl-C₁₋₆ alkyl, C₁₋₆ alkoxy substituted phenyl-C₁₋₆alkyl, C₁₋₆ alkyl, amino, carboxy, C₁₋₆ alkylamino, C₁₋₆ alkoxycarbonyl,di(C₁₋₆ alkyl)amino, benzylamino, C₁₋₆ alkylsulfonyl, piperidino C₁₋₆alkyl carbonyl, or optionally fused by benzene; or

—NR³⁴R³⁵,

 wherein R³⁴ is hydrogen and

R³⁵ is —(CH₂)_(m)—NR³⁵¹R³⁵² (m represents any of integers from 1 to 6)

wherein R³⁵¹ represents hydrogen, C₁₋₆ alkyl,

R³⁵² represents hydrogen, C₁₋₆ alkyl, C₁₋₆ alkanoyl, C₁₋₆alkylsubstituted phenyl, benzoyl, C₁₋₆ alkanoyl, phenylaminocarbonyl,phenylsulfonyl; and

R⁴ represents cyano, C₁₋₆ alkyl optionally substituted by hydroxy orC₁₋₆ alkoxy, or

—(CH₂)_(p)NHCOR⁴¹, —(CH₂)_(p)NHC(═S)R⁴¹

 wherein p represents any of integer from 1 to 6 and R⁴¹ represents C₁₋₆alkoxy, amino, phenylamino, C₁₋₆ alkyl, C₁₋₆ alkylamino, di(C₁₋₆alkyl)amino, C₃₋₁₀ cycloalkylamino;

R⁵ represents amino, or

R⁴ and R⁵ may form,

—R⁴⁰—CO—NH—, —R⁴⁰—SO₂—NH—,

—R⁴⁰—C(═S)—NH— or

—R⁴⁰—CH₂—NH—,

 wherein said —R⁴⁰— represents —CHR⁴⁰¹—O—, —CH₂—NR⁴⁰¹—, —CO—NR⁴⁰¹—, (inwhich R⁴⁰¹ is hydrogen, C₁₋₆ alkanoyl, C₁₋₆ alkoxycarbonyl, C₁₋₆ alkyl,phenyl, C₁₋₆ alkylsulfonyl, C₃₋₈ cycloalkylaminocarbonyl, C₁₋₆alkylaminocarbonyl, carbamoyl, di(C₁₋₆ alkyl)aminocarbonyl),—CH₂CHR⁴⁰²—, —CH═CR⁴⁰²—, (in which R⁴⁰² is hydrogen, halogen, nitro,amino, ciano, benzoylamino, phenylsulfonyl, carbamoyl, hydroxy-carbonyl,C₁₋₆ alkoxycarbonyl, C₁₋₁₂ alkylaminocarbonyl, halogen substituted C₁₋₆alkylaminocarbonyl, C₁₋₆ alkanoylamino, C₁₋₆ alkylamino, di(C₁₋₆alkyl)aminocarbonyl, di(C₁₋₆ alkyl)aminoC₁₋₆ alkylaminocarbonyl,hydroindenylaminocarbonyl, diphenylmethylaminocarbonyl,pyrrolidinocarbonyl, C₁₋₆ alkoxy C₁₋₆ alkyl amino carbonyl,morpholinocarbonyl, piperazinocarbonyl, phenylC₁₋₆ alkylaminocarbonyl,C₃₋₈ cycloalkylaminocarbonyl, hydroxycarbonylC₁₋₆ alkylaminocarbonyl,C₃₋₈ cycloalkylC₁₋₆ alkylaminocarbonyl, hydroxyC₁₋₆ alkylaminocarbonyl,carboxyethylaminocarbonyl, methylsulfonylaminocarbonyl,)

—CR⁴¹═N—NH— (R⁴¹ is hydroxy, amino, C₁₋₆ alkanoylamino) or

—CR⁴²═N—C═N— (R⁴² is amino)

or a salt thereof.

The preferable compounds of the present invention are as follows or thesalt thereof:

7-(2-hydroxyphenyl)-5-(3-piperidinyl)-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-2-one;

2-amino-6-[2-(benzyloxy)-6-hydroxyphenyl]-4-(3-piperidinyl)nicotinonitrile;

2-amino-6-(2-hydroxy-6-propoxyphenyl)-4-(3-piperidinyl)nicotinonitrile;

2-[6-amino-5-(hydroxymethyl)-4-(3-piperidinyl)-2-pyridinyl]-3-(benzyloxy)phenol;

7-[2-(benzyloxy)-6-hydroxyphenyl]-5-(3-piperidinyl)-1,4-dihydro-2H-pyrido[2,3d][1,3]oxazin-2-one;

2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-(3-piperidinyl)nicotinonitriletrifluoroacetate;

7-(2-hydroxyphenyl)-5-(3-piperidinyl)-3,4-dihydro-1,8-naphthyridin-2(1H)-one;

2-amino-6-[2-(cyclobutylmethoxy)-6-hydroxyphenyl]-4-(3-piperidinyl)nicotinonitrile;

2-[6-amino-5-(hydroxymethyl)-4-(3-piperidinyl)-2-pyridinyl]-3-propoxyphenol;

7-(2-hydroxy-6-propoxyphenyl)-5-(3-piperidinyl)-1,4-dihydro-2H-pyrido[2,3d][1,3]-oxazin-2-one;

ethyl7-(2-hydroxy-6-propoxyphenyl)-2-oxo-5-(3-piperidinyl)-1,2,3,4-tetrahydro-1,8-naphthyridine-3-carboxylate;

7-(2-hydroxy-6-propoxyphenyl)-5-(3-piperidinyl)-3,4-dihydro-1,8-naphthyridin-2(1H)-one;

2-[6-amino-5-(hydroxymethyl)-4-(3-piperidinyl)-2-pyridinyl]-3-(cyclopropylmethoxy)phenol;

2-[6-amino-5-(hydroxymethyl)-4-(4-piperidinyl)-2-pyridinyl]-3-(cyclopropylmethoxy)phenol;

7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-5-(3-piperidinyl)-3,4-dihydro-1,8-naphthyridin-2(1H)-one;

ethyl7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-5-(3-piperidinyl)-1,2,3,4-tetrahydro-1,8-naphthyridine-3-carboxylate;

7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-5-(4-piperidinyl)-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-2-one;6′-amino-5′-(hydroxymethyl)-4′-(3-piperidinyl)-2,2′-bipyridin-3-ol;

7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-5-(4-piperidinyl)-3,4-dihydro-1,8-naphthyridin-2(1H)-one;

7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-3-fluoro-5-(3-piperidinyl)-1,8-naphthyridin-2(1H)-one;

7-(2-hydroxy-6-propoxyphenyl)-5-(4-piperidinyl)-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-2-one;

7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-5-(3-piperidinyl)-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-2-one;

3-(cyclopropylmethoxy)-2-[5-(3-piperidinyl)-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-7-yl]phenol;

2-[6-amino-5-(hydroxymethyl)-4-(3-piperidinyl)-2-pyridinyl]-3-(neopentyloxy)phenol;

2-[6′-amino-5′-(hydroxymethyl)-1,2,5,6-tetrahydro-3,4′-bipyridin-2′-yl]phenol;

7-[2-(cyclopropyhethoxy)-6-hydroxyphenyl]-5-(3-piperidinyl)-1,8-naphthyridin-2(1H)-one;

N-{[2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-(3-piperidinyl)-3-pyridinyl]methyl}acetamide;

7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-5-(3-piperidinyl)-1,2-dihydro-1,8-naphthyridine-3-carboxamide;

3-acetyl-7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-5-(3-piperidinyl)-3,4-dihydropyrido[2,3-d]pyrimidin-2(1H)-one;

2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-(4-piperidinyl)nicotinonitrile;

2-amino-4-[(2-aminoethyl)amino]-6-[2-(cyclopropylmethoxy)-6-hydroxy-phenyl]nicotinonitrile;

N-{[2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-(3-piperidinyl)-3-pyridinyl]methyl}-N′-propylurea;

7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-5-(3-piperidinyl)-3,4-dihydro-pyrido[2,3-d]pyrimidin-2(1H)-one;

ethyl[2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-(3-piperidinyl)-3-pyridinyl]methylcarbamate;

2-amino-6-{2-hydroxy-6-[(4-methylpentyl)oxy]phenyl}-4-(4-piperidinyl)nicotinonitrile;

7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-5-(3-piperidinyl)-1,2,3,4-tetrahydro-1,8-naphthyridine-3-carboxamide;

7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-N-isopropyl-2-oxo-5-(3-piperidinyl)-1,2-dihydro-1,8-naphthyridine-3-carboxamide;

ethyl7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-5-(3-piperidinyl)-1,4-dihydropyrido[2,3-d]pyrimidine-3(2H)-carboxylate;

N-{[2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-(3-piperidinyl)-3-pyridinyl]methyl}urea;

2-amino-6-(2-hydroxy-6-propoxyphenyl)-4-(4-piperidinyl)nicotinonitrile;

N-cyclohexyl-7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-5-(3-piperidinyl)-1,4-dihydropyrido[2,3-d]pyrimidine-3(2H)-carboxamide;

2-amino-6-[2-(cyclobutylmethoxy)-6-hydroxyphenyl]-4-(4-piperidinyl)nicotinonitrile;

7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-N,N-dimethyl-2-oxo-5-(3-piperidinyl)-1,4-dihydropyrido[2,3-d]pyrimidine-3(2H)-carboxamide;

2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-(1-methyl-3-piperidinyl)nicotinonitrile;

7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-(3-piperidinyl)-1,4-dihydro-pyrido[2,3-d]pyrimidine-3(2H)-carboxamide;

isopropyl[2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-(3-piperidinyl)-3-pyridinyl]methylcarbamate;

isopropyl7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-5-(3-piperidinyl)-1,4-dihydropyrido[2,3-d]pyrimidine-3(2H)-carboxylate;

isobutyl7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-5-(3-piperidinyl)-1,4-dihydropyrido[2,3-d]pyrimidine-3(2H)-carboxylate;

neopentyl7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-5-(3-piperidinyl)-1,4-dihydropyrido[2,3-d]pyrimidine-3(2H)-carboxylate;neopentyl[2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-(3-piperidinyl)-3-pyridinyl]methylcarbamate;

2-amino-6-[2-(hexyloxy)-6-hydroxyphenyl]-4-(4-piperidinyl)nicotinonitrile;and

7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-N-ethyl-2-oxo-5-(3-piperidinyl)-1,4-dihydropyrido[2,3-d]pyrimidine-3(2H)-carboxamide

The compound of the formula (I) of the present invention can be, but notlimited to be, prepared by combining various known methods. In someembodiments, one or more of the substituents, such as amino group,carboxyl group, and hydroxyl group of the compounds used as startingmaterials or intermediates are advantageously protected by a protectinggroup known to those skilled in the art. Examples of the protectinggroups are described in “Protective Groups in Organic Synthesis (2^(nd)Edition)” by Greene and Wuts.

The compound (I-a)

wherein X is CH or N, R¹¹, R², and R³ are the same as defined,

or a salt thereof, can be prepared, for example, by the followingreaction A.

The compound of the formula (II)

in which R¹¹ and R² are the same as defined above,

are reacted with an aldehyde of the formula R³—CHO(III), a nitrile ofthe formula NCCH₂CN (IV), and an ammonium salt such as ammonium acetate.R³ is the same as defined above. R³′—CHO(III′) can be advantageouslyused instead of R³—CHO(III) in some instances. R³′ can representesterified R³ by ethyl, tertiary butyl or the like: or other esters orother substituents which can be easily converted to R³ by conventionalmethods. Hydroxyl group of the compound of the formula (II) is protectedby an appropriate protecting group (e.g., benzyl, methoxybenzy, andsilyl) during the reaction, and deprotected afterward. R³′ can also betreated by acids to obtain R³.

In the sketch above, CN at position C-3 can be replaced by carboxylatesderived from tertiary alcohol, such as COOtBu with the use ofNCCH₂—COOtBu (IV′) instead of NCCH₂CN (IV). In this case the followingcompound (I-a′) can be obtained.

The step 1 and step 1′ of the reaction A can be carried out without asolvent or in a solvent including, for instance, ethers, such asdioxane, and tetrahydrofuran; aromatic hydrocarbons such as benzene,toluene and xylene; nitrites such as acetonitrile; amides such asdimethylformamide (DMF) and dimethylacetamide; sulfoxides such asdimethyl sulfoxide, and others.

The reaction temperature can be optionally set depending on thecompounds to be reacted. The reaction temperature is usually, but notlimited to, about 50° C. to 200° C. The reaction may be conducted for,usually, 30 minutes to 48 hours and preferably 1 to 24 hours.

The compounds of the general formula (II), (III), (III′) can becommercially available, or can be prepared by the use of knowntechniques.

Step 2 and step 2′ of the reaction A can be carried out for example,under the hydrogen atmosphere with hydrogeneous catalysis, such as Pd-Cin a solvent including, for instance, esters, such as ethyl acetate,ethers, such as dioxane, and tetrahydrofuran; aromatic hydrocarbons suchas benzene, toluene and xylene; nitrites such as acetonitrile; amidessuch as dimethylformamide (DMF) and dimethylacetamide; sulfoxides suchas dimethyl sulfoxide, and others.

The reaction temperature can be, but not limited to, about 50° C. to200° C. The reaction may be conducted for, usually, 30 minutes to 48hours and preferably 1 to 24 hours.

Step 3 and Step 3′ of the reaction A can be any kind of conventionalreaction starting from ester to obtain R³, e.g., acid treatment, alkalitreatment, amidation, and hydrogenation: or other reaction such asalkylation or the like to obtain R³.

Alternatively, the compound (I-b)

wherein X, R², and R³ are the same as defined above and Y is C₁₋₁₂ alkylor R¹¹¹—(CH₂)_(n)—, in which R¹¹¹ and n are the same as defined above,or a salt thereof can be obtained by the following reaction B.

The compound (I-b′) can be also obtained in the reaction B.

In reaction B, the compound of the formula (II-a) is reacted with analdehyde (III), a nitrile (IV) and ammonium acetate under the samecondition as the reaction A to obtain the compound of the formula (V′).The benzyl protecting groups in the compound of the general formula(II-a) can be replaced with any of appropriate protecting group. Theprotecting group is then removed after the reaction. In the step 4 ofthe reaction B, the compound (V) is reacted with L-Y, wherein Lrepresents a leaving group, such as halogen atom e.g., chlorine, bromineor iodine atom; C_(6-C) ₁₀ arylsulfonyloxy group e.g.benzenesulfonyloxy, polysulfonyloxy, or p-toluene-sulfonyloxy; andC_(1-C) ₄ alkylsulfonyloxy group, e.g. methanesulfonyloxy and the like.Y represents C_(1-C) ₆ alkyl, or —(CH₂)_(n)—R¹¹¹ (wherein R¹¹¹ is thesame as defined above). The reaction with the compound (V) and L—Y canbe carried out in a solvent including, for instance, alcohols such asmethanol and ethanol; ethers, such as dioxane, and tetrahydrofuran(THF); nitrites such as acetonitrile; amides such as dimethylformamide(DMF) and dimethylacetamide; sulfoxides such as dimethyl sulfoxide, andothers. Optionally, two or more of the solvents selected from the listedabove can be mixed and used.

The reaction temperature of the reaction between compound (V) and L—Ycan be optionally set depending on the compounds to be reacted. Thereaction temperature is usually, but not limited to, about −10° C. to200° C. and preferably about 10° C. to 80° C. The reaction may becarried out for, usually, 30 minutes to 48 hrs and preferably 1 to 24hrs. The reaction can be advantageously conducted in the presence of abase. Examples of the base include an alkali metal hydride such assodium hydride or potassium hydride; alkali metal alkoxide such assodium methoxide or sodium ethoxide; alkali metal hydroxide such assodium hydroxide or potassium hydroxide; carbonates such as sodiumcarbonate or potassium carbonate, and hydrogen carbonates such as sodiumhydrogen carbonate and potassium hydrogen carbonate; organic amines suchas triethylamine.

The step 3 of the reaction B is the same as that of reaction A.

Alternatively, the compound of the formula (I-c) below:

wherein X, R¹¹, R², R³⁴ and R³⁵ are the same as defined above, can beadvantageously prepared by the following reaction C.

First, the compound of the formula (VI) may be reacted with carbondisulfide and R⁶⁰—L (wherein R⁶⁰ represents C₁₋₆ alkyl and L representsa leaving group as defined above )to obtain the compound of the formula(VII). Benzyl protecting group in the compound of the formula (VI) canbe replaced with any of an appropriate protecting group. This reactionmay be advantageously conducted in the presence of base, such as thecombination of sodium hydride and dimethyl acetamide.

The resulting compound (VII) may be reacted with cyanoacetamide in thepresence of a solvent and a base. Then the compound (VIII) may beoxidized to yield the compound (IX). The compound (IX) is then reactedwith halogenoacetoamide such as chloroacetamide in the presence of abase in a solvent. The resulting compound (X) is reacted with NHR³⁴R³⁵(and R³⁵ are the same as defined above). Finally, the generated productis reacted with a base and is deprotected to obtain the compound (I-c).

The solvents used in each process of the reaction include, for instance,ethers, such as dioxane and tetrahydrofuran; aromatic hydrocarbons suchas benzene, toluene and xylene; nitrites such as acetonitrile; amidessuch as dimethylformaide (DMF) and dimethylacetamide; sulfoxides such asdimethyl sulfoxide, and others. The above solvent may be used alone orin combination.

Examples of the base used in the reaction include an alkali metalhydride such as sodium hydride or potassium hydride; alkali metalalkoxide such as sodium methoxide or sodium ethoxide; alkali metalhydroxide such as sodium hydroxide or potassium hydroxide; carbonatessuch as sodium carbonate or potassium carbonate, and hydrogen carbonatessuch as sodium hydrogen carbonate and potassium hydrogen carbonate;organic amines such as triethylamine.

The reaction temperature can be optionally set depending on the compoundto be reacted. The reaction temperature, unless otherwise stated above,is about 10° C. to 200° C. Each process of the reaction may be conductedfor, usually, 30 minutes to 48 hours and preferably 1 to 24 hours.

Amino group at position 2 of the pyridine ring is, if necessary,modified according to conventional method to prepare other groups suchas alkylamino, alkanoylamino, etc.

When R¹¹ is C₁₋₆ alkylsulfonylamino, C₁₋₆ alkylamino, di(C₁₋₆alkyl)amino, C₁₋₆ alkanoylamino, phenyl C₁₋₆ alkylamino, orphenylsulfonylamino, it is derived from —NH₂ with the use ofconventional methods during the course of reaction A.

The compounds of the formulas (I-a), (I-b) and (I-c) can be furtherreacted to modify the substituents at position 2 and position 3 of thepyridine ring to synthesize the desired compounds in the scope of thepresent invention. Also, in the course of reaction A, B, and C above,the substituents at position 2 and position 3 of the pyridine ring canbe modified.

The amino moiety at position 2 can be modified by the conventionalmethods as follows:

wherein L is the leaving group and the same as defined above, Z isbenzyl, C₁₋₆ alkyl, or phenyl, or NHZ may form saturated 5-6 memberedring optionally contain NH or O as other heteroatom than the adjacent Natom.

In another embodiment, the amino moiety at position 2 can be convertedto amide with the use of acid chloride.

The cyano moiety at position 3 can be converted to carbamoyl by theconventional alkaline hydrolysis.

The tertiary butoxy carbonyl at position 3 can be easily modified, bythe conventional reactions of ester, to alcohol, carboxyl, and the like.Alcohol or carboxy may be further converted to another substituents bythe conventional methods.

In some embodiment, the substituents of the positions 2 and 3 togetherform ring optionally having substituents. Any conventional method orcombination of any conventional methods can be used to form the rings.The examples of forming the rings are shown below.

Yet, in another embodiment, the substituents of the positions 3 and 4together form ring optionally having substituents. Any conventionalmethod or combination of any conventional methods can be used to formthe rings. The examples of forming the rings are shown below.

When the compound shown by the formula (I) or a salt thereof hastautomeric isomers and/or stereoisomers (e.g, geometrical isomers andconformational isomers), each of their separated isomer and mixtures arealso included in the scope of the present invention.

When the compound shown by the formula (I) or a salt thereof has anasymmetric carbon in the structure, their optically active compounds andracemic mixtures are also included in the scope of the presentinvention.

Typical salts of the compound shown by the formula (I) include saltsprepared by reaction of the compounds of the present invention with amineral or organic acid, or an organic or inorganic base. Such salts areknown as acid addition and base addition salts, respectively.

Acids to form acid addition salts include inorganic acids such as,without limitation, sulfuric acid, phosphoric acid, hydrochloric acid,hydrobromic acid, hydriodic acid and the like, and organic acids, suchas, without limitation, p-toluenesulfonic acid, methanesulfonic acid,oxalic acid, p-bromophenylsulfonic acid, carbonic acid, succinic acid,citric acid, benzoic acid, acetic acid, and the like.

Base addition salts include those derived from inorganic bases, such as,without limitation, ammonium hydroxide, alkaline metal hydroxide,alkaline earth metal hydroxides, carbonates, bicarbonates, and the like,and organic bases, such as, without limitation, ethanolamine,triethylamine, tris(hydroxymethyl)aminomethane, and the like. Examplesof inorganic bases include, sodium hydroxide, potassium hydroxide,potassium carbonate, sodium carbonate, sodium bicarbonate, potassiumbicarbonate, calcium hydroxide, calcium carbonate, and the like.

The compound of the present invention or a salts thereof, depending onits substituents, may be modified to form lower alkylesters or knownother esters; and/or hydrates or other solvates. Those esters, hydrates,and solvates are included in the scope of the present invention.

The compound of the present invention may be administered in oral forms,such as, without limitation normal and enteric coated tablets, capsules,pills, powders, granules, elixirs, tinctures, solution, suspensions,syrups, solid and liquid aerosols and emulsions. They may also beadministered in parenteral forms, such as, without limitation,intravenous, intraperitoneal, subcutaneous, intramuscular, and the likeforms, well known to those of ordinary skill in the pharmaceutical arts.The compounds of the present invention can be administered in intranasalform via topical use of suitable intranasal vehicles, or via transdermalroutes, using transdermal delivery systems well known to those ofordinary skilled in the art.

The dosage regimen with the use of the compounds of the presentinvention is selected by one of ordinary skill in the arts, in view of avariety of factors, including, without limitation, age, weight, sex, andmedical condition of the recipient, the severity of the condition to betreated, the route of administration, the level of metabolic andexcretory function of the recipient, the dosage form employed, theparticular compound and salt thereof employed.

The compounds of the present invention are preferably formulated priorto administration together with one or more pharmaceutically acceptableexcipients. Excipients are inert substances such as, without limitationcarriers, diluents, flavoring agents, sweeteners, lubricants,solubilizers, suspending agents, binders, tablet disintegrating agentsand encapsulating material.

Yet, another embodiment of the present invention is pharmaceuticalformulation comprising a compound of the invention and one or morepharmaceutically acceptable excipients that are compatible with theother ingredients of the formulation and not deleterious to therecipient thereof. Pharmaceutical formulations of the invention areprepared by combining a therapeutically effective amount of thecompounds of the invention together with one or more pharmaceuticallyacceptable excipients therefor. In making the compositions of thepresent invention, the active ingredient may be mixed with a diluent, orenclosed within a carrier, which may be in the form of a capsule,sachet, paper, or other container. The carrier may serve as a diluent,which may be solid, semi-solid, or liquid material which acts as avehicle, or can be in the form of tablets, pills powders, lozenges,elixirs, suspensions, emulsions, solutions, syrups, aerosols, ointments,containing, for example, up to 10% by weight of the active compound,soft and hard gelatin capsules, suppositories, sterile injectablesolutions and sterile packaged powders.

For oral administration, the active ingredient may be combined with anoral, and non-toxic, pharmaceutically-acceptable carrier, such as,without limitation, lactose, starch, sucrose, glucose, sodium carbonate,mannitol, sorbitol, calcium carbonate, calcium phosphate, calciumsulfate, methyl cellulose, and the like; together with, optionally,disintegrating agents, such as, without limitation, maize, starch,methyl cellulose, agar bentonite, xanthan gum, alginic acid, and thelike; and optionally, binding agents, for example, without limitation,gelatin, acacia, natural sugars, beta-lactose, corn sweeteners, naturaland synthetic gums, acacia, tragacanth, sodium alginate,carboxymethylcellulose, polyethylene glycol, waxes, and the like; and,optionally, lubricating agents, for example, without limitation,magnesium stearate, sodium stearate, stearic acid, sodium oleate, sodiumbenzoate, sodium acetate, sodium chloride, talc, and the like.

In powder forms, the carrier may be a finely divided solid, which is inadmixture with the finely divided active ingredient. The activeingredient may be mixed with a carrier having binding properties insuitable proportions and compacted in the shape and size desired toproduce tablets. The powders and tablets preferably contain from about 1to about 99 weight percent of the active ingredient which is the novelcomposition of the present invention. Suitable solid carriers aremagnesium carboxymethyl cellulose, low melting waxes, and cocoa butter.

Sterile liquid formulations include suspensions, emulsions, syrups andelixirs. The active ingredient can be dissolved or suspended in apharmaceutically acceptable carrier, such as sterile water, sterileorganic solvent, or a mixture of both sterile water and sterile organicsolvent.

The active ingredient can also be dissolved in a suitable organicsolvent, for example, aqueous propylene glycol. Other compositions canbe made by dispersing the finely divided active ingredient in aqueousstarch or sodium carboxymethyl cellulose solution or in suitable oil.

The formulation may be in unit dosage form, which is a physicallydiscrete unit containing a unit dose, suitable for administration inhuman or other mammals. A unit dosage form can be a capsule or tablets,or a number of capsules or tablets. A “unit dose” is a predeterminedquantity of the active compound of the present invention, calculated toproduce the desired therapeutic effect, in association with one or moreexcipients. The quantity of active ingredient in a unit dose may bevaried or adjusted from about 0.1 to about 1000 milligrams or moreaccording to the particular treatment involved.

Typical oral dosages of the present invention, when used for theindicated effects, will range from about 0.01 mg/kg/day to about 100mg/kg/day, preferably from 0.1 mg/kg/day to 30 mg/kg/day, and mostpreferably from about 0.5 mg/kg/day to about 10 mg/kg/day. In the caseof parenteral administration, it has generally proven advantageous toadminister quantities of about 0.001 to 100 mg/kg/day, preferably from0.01 mg/kg/day to 1 mg/kg/day. The compounds of the present inventionmay be administered in a single daily dose, or the total daily dose maybe administered in divided doses, two, three, or more times per day.Where delivery is via transdermal forms, of course, administration iscontinuous.

The effect of the present compounds was examined by the following assaysand pharmacological tests.

IKK-β Kinase Inhibitory Assay

(1) Preparation of IKK-β Kinase Protein.

A cDNA fragment encoding human IKK-β open reading frame was generated byPCR with the use of a pair of primers designed from the publishedsequence (Woronicz J D et al. (1997) Science 278, 866-869). A templatewas obtained from Quickclone cDNA (Clontech) using Elongase™Amplification kit (Life Technologies). The DNA fragments generated byPCR were gel-purified and subcloned into pBluescript. The cDNA fragmentcloned in pBluescript was inserted into pcDNA3.1/His C KpnI/Notl, andtransferred into pVL1393 SmaI/XbaI (Pharmingen) to construct abaculovirus transfer vector. Then the vector, together with thelinearized baculovirus (BaculoGold™, Pharmingen) was used to transfectSf21 cells (Invitrogen, San Diego, Calif.). Generated recombinantbaculovirus was cloned and amplified in Sf21 cells, grown in TNM-FHinsect cell medium (Life Technologies, Inc.) supplemented with 10% FCS,50 g/ml Gentamycin, 0.1% Pluronic F-68 (Life Technologies, Inc.) assuspension culture (200 ml in 1 L Erlenmeyer flask; 27° C.; 130 rpm).Sf21 cells were infected with this amplified virus with a multiplicityof infection of 5 following standard protocols (Crossen R, Gruenwald S(1997) Baculovirus Expression Vector System Instruction Manual,Pharmingen Corporation) and harvested 48 hrs later. The cells were lysedto obtain the produced chimeric protein of IKK-β kinase fused byhistidine (His-tagged IKK-beta).

(2) The Preparation of Purified GST-IκBα Fusion Proteins

An expression vector containing the nucleotide sequence encoding fusionprotein of GST with amino acid residues 1 to 54 of IκBα under thecontrol of an IPTG-inducible promoter was constructed. The expressionvector was introduced in E. coli and the transformant was cultured andlysed to obtain a GST-IκBα fusion protein. Then the resulting GST-IκBαfusion protein was purified and biotinated for kinase assay.

(3) The Measurement of IKK-β Kinase Activity

The 96-well format kinase assay of IKK-β were performed to test theinhibitory activity of the compounds of the present invention. First, 5μl of a test compound was put in the presence of 2.5% dimethyl sulfoxide(DMSO) in each well in a U-bottomed 96-well plate (Falcon). For controlwells of background (BG) and total phosphorylation (TP), 5 μl of 2.5%DMSO was put. Recombinant IKK-β (final 0.6 μg/ml) and bio-GST-IκBα(1-54) (final 0.2 μM) were diluted in 25 μl of 2×kinase buffer β (40 mMTris-HCl, pH 7.6, 40 mM MgCl₂, 40 mM β-glycerophosphate, 40 mMp-nitro-phenylphosphate, 2 mM EDTA, 40 mM creatine phosphate, 2 mM DTT,2 mM Na₃VO₄, 0.2 mg/ml BSA and 0.8 mM phenylmethylsulfonyl fluoride) andtransferred to the 96-well plate. Bio-GST-IκBα (1-54) in 25 μl of2×kinase buffer β without IKK-β was transferred to BG wells. Then 20 μlof 12.5 μM ATP, 62.5 μCi/ml [γ-³³P] ATP (Amersham Pharmacia Biotech) wasadded and the resulting mixture was incubated for 2 hrs at roomtemperature. The kinase reactions were terminated by the addition of 150μl of termination buffer (100 mM EDTA, 1 mg/ml BSA, 0.2 mg NaN₃). Onehandred and fifty μl of the sample were transferred to astreptavidin-coated, white MTP (Steffens Biotechniche Analysen GmbH#08114E14.FWD) to capture the biotinylated substrates. After 1 hr ofincubation, non-bound radioactivity was eliminated by washing the wellsfive times with 300 μl of washing buffer including 0.9% NaCl and 0.1%(w/v) Tween-20 with the use of a MW-96 plate washer (BioTec). The boundradioactivity was determined after the addition of 170 μl MicroScint-PSscintillation cocktail (Packard) using a TopCount scintillation counter.

Syk Tyrosine Kinase Inhibitory Assay for Selectivity

(1) Preparation of Syk Protein

A cDNA fragment encoding human Syk openreading frame was cloned fromtotal RNA of human Burkitt's lymphoma B cell lines, Raji (American TypeCulture Collection), with the use of RT-PCR method. The cDNA fragmentwas inserted into pAcG2T (Pharmingen, San Diego, Calif.) to construct abaculovirus transfer vector. Then the vector, together with thelinearized baculovirus (BaculoGold™, Pharmingen), was used to transfectSf21 cells (Invitrogen, San Diego, Calif.).

Generated recombinant baculovirus was cloned and amplified in Sf21cells. Sf21 cells were infected with this amplified high titer virus toproduce a chimeric protein of Syk kinase fused byglutathione-S-transferase (GST).

The resulting GST-Syk was purified with the use of glutathione column(Amersham Pharmacia Biotech AB, Uppsala, Sweden) according to themanufacturer's instruction. The purity of the protein was confirmed tobe more than 90% by SDS-PAGE.

(2) Synthesize of a Peptide

Next, a peptide fragment of 30 residues including two tyrosine residues,KISDFGLSKALRADENYYKAQTHGKWPVKW, was synthesized by a peptidesynthesizer. The N-terminal of the fragment was then biotinylated toobtain biotinylated activation loop peptide (AL).

(3) The Measurement of Syk Tyrosine Kinase Activity

All reagents were diluted with the Syk kinase assay buffer (50 mMTris-HCl (pH 8.0), 10 mM MgCl₂, 0.1 mM Na₃VO₄, 0.1% BSA, 1 mM DTT).First, a mixture (35 μl) including 3.2 μg of GST-Syk and 0.5 μg of ALwas put in each well in 96-well plates. Then 5 μl of a test compound inthe presence of 2.5% dimethyl sulfoxide (DMSO) was added to each well.To this mixture was added 300 μM ATP (10 μl) to initiate the kinasereaction. The final reaction mixture (50 μl) consists of 0.65 nMGST-Syk, 3 μM AL, 30 μM ATP, a test compound, 0.25% DMSO, and a Sykkinase assay buffer.

The mixture was incubated for 1 hr at room temperature (RT), and thereaction was terminated by the addition of 120 μl of termination buffer(50 mM Tris-HCl (pH 8.0), 10 mM EDTA, 500 mM NaCl, 0.1% BSA). Themixture was transferred to streptavidin-coated plates and incubated for30 min. at room temperature to combine biotin-AL to the plates. Afterwashing the plates with Tris-buffered saline (TBS) (50 mM Tris-HCl (H8.0), 138 mM NaCl, 2.7 mM KCl) containing 0.05% Tween-20 for 3 times,100 μl of antibody solution consisting of 50 mM Tris-HCl (pH 8.0), 138mM NaCl, 2.7 mM KCl, 1% BSA, 60 ng/ml anti-phosphotyrosine monoclonalantibody, 4G10 (Upstate Biotechnology), which was labeled with europiumby Amersham Pharmacia's kit in advance, was added and incubated at roomtemperature for 60 minutes. After washing, 100 μl of enhancementsolution (Amersham Pharmacia Biotech) was added and then time-resolvedfluorescence was measured by multi-label counter ARVO (Wallac Oy,Finland) at 340 nm for excitation and 615 nm for emission with 400 msecof delay and 400 msec of window.

The Measurement of RANTES Production in Response to TNF-α from A549Cells

(1) Preparation of A549 Cells

The A549 human lung epithelium cell line (ATCC #CCL-885) was maintainedin Dulbecco's modified Eagle's medium (D-MEM, Nikken BiomedicalInstitute) supplemented with 10% FCS (Gibco), 100 U/ml penicillin, 100μg/ml streptomycin, and 2 mM glutamine (culture medium). Forty thousand(4×10⁴) cells (80 μl/well) were seeded in each well of 96 wellflat-bottom tissue culture plate (Falcon #3072). The plate was allowedto stand for 2 hrs, thus the cells were adhered to the bottom of eachwell. To the each well was added 10 μl vehicle (1% DMSO), serialdilutions of test compounds in 1% DMSO, or 5 nM Dexamethasone in 1% DMSOas a reference. The mixture (90 μl/well) was incubated for 1 hr at 37°C. After 1 hr, 1 μg/ml TNF-α (10 μl) in culture medium was added to themixture to obtain 100 μl of reaction mixture. The reaction mixture wascultured for 24 hrs to stimulate the cells with 100 ng/ml TNF-α. Cellswith vehicle without TNF-α stimulation were also prepared.

(2) Measurement of RANTES Production

Then the concentration of RANTES released from the cells in thesupernatants of each well was determined using a quantitative sandwichenzyme immunoassay technique. First, 2 μg/ml mouse anti-huRANTES mAb(R&D Systems, #mAb678) in PBS buffer (pH 7.4, 100 μl) was put in eachwell of 96-well NUNC fluoro plate (Nalge Nunc, New York USA) (Final 200ng/well) and the plate was allowed to stand for overnight at 4° C. to becoated by the antibody. Each well of the plate was then washed with 350μl wash buffer (0.05% Tween-20, 0.85% NaCl, and 25 mM Tris/HCl pH 7.4)for three times. Blocking buffer containing 1% BSA (Sigma 99% pure, 100g), 5% sucrose (Nacalai tesque, 99% pure, 500 g), and 0.02% azide(Nacalai tesque, 100%, 500 g) were added (200 μl) to each well and thenthe plate was allowed to stand for 4 hours to stabilize the coatedantibody. Next, 50 μl supernatants of cell culture prepared in (1) abovewere put in each well of the 96-well NUNC fluoro plate with coatedantibody. Recombinant Human RANTES (Pepro Tech, Inc. #300-06) was usedas the standard for the determination of RANTES production (linear rangebetween 1 and 10 ng/ml). Eu-labelled mouse anti-huRANES mAb (60 ng/ml:R&D Systems, #mAb278) in PBS supplemented by 1% BSA and 0.05% Tween 20was added (50 μl) to each well. The reaction mixtures were incubated atroom temperature for 4 hrs. After washing with wash buffer (0.05%Tween-20, 0.85% NaCl, and 25 mM Tris/HCl pH 7.4, 350 μl/well) for 5times with the use of a Sera Washer (Bio-Tech, #MW-96R), the enhancementsolution (DELFIA, #1244-405, 100 μl/well) was added to each well. Theplate was incubated for 10 minutes at room temperature with moderateshaking. Fluorescent intensity was measured using a DELFIA fluorimeter(Wallac). Excitation was performed at 340 nm and emission was measuredat 615 nm.

The Measurement of TNF-α Production in Response to LPS from PeripheralBlood Mononuclear Cells (PBMC)

(1) Preparation of PBMC

Human PBMC were prepared by first obtaining blood from healthy donorsand isolating the cells from the blood. The isolation was done by Ficollgradient-centrifugation method using Ficoll Pacque (Pharmacia#17-1440-02). Within three hours from donation, the isolated PBMC wasused. After three times washing with PBS, PBMC were resuspended withRPMI 1640 (Nikken BioMedical Institute) supplemented with 10% FCS(Gibco), 100 U/ml penicillin, 100 μg/ml streptomycin, and 2 mM glutamine(culture medium). The cells (1×10⁵ in 150 μl/well) were seeded in eachwell of 96 well flat-bottom tissue culture plate (Falcon #3072). To theeach well was added 20 μl vehicle (1% DMSO), serial dilutions of testcompounds in 1% DMSO, or 250 nM Dexamethasone in 1% DMSO as a reference.The mixture (170 μl/well) was incubated for 1 hr at 37° C. After 1 hr,20 ng/ml LPS (30 μl) in culture medium was added to the mixture toobtain 200 μl of reaction mixture. The reaction mixture was cultured for7 hrs to stimulate the cells with 3 ng/ml LPS. Cells with vehiclewithout LPS stimulation were also prepared. The supernatants of thereaction mixture were then collected.

(2) Measurement of TNF-α Production

The TNF-α concentration in the supernatants was determined using aDuoSet™ ELISA Development Kit (GenzymeTechne, Minneapolis, USA)following the manufacturer's recommendations. First, 4 μg/ml of mouseanti-human TNF-α Ab in PBS buffer (100 μl) was put in each well of96-well plate (NUNC, Maxisorp™) and the plate was allowed to stand forovernight at 4° C. to be coated with the antibody. Each well of theplate was then washed 5 times with 350 μl of wash buffer containing PBS,0.05% Tween 20 (Nakalai tesque) using Sera Washer (Bio-Tech, #MW-96R).To each well was added 300 μl of 1% BSA (Sigma), 5% sucrose in PBS.After 2 hrs incubation at room temperature, the buffer was discarded,and 50 μl of culture medium was added. Next, 50 μl supernatant ofstimulated cell culture prepared (1) above was put in each well of the96-well plate. Recombinant human TNF-α (Genzyme Techne) was used as thestandard for the determination of TNF-α production (linear range between30 and 2,000 μg/ml). The reaction mixtures were incubated for 1 hr atroom temperature. After 5 times washing, 100 μl biotinylated goatanti-human TNF-α antibody (Genzyme Techne, 300 ng/ml) in 0.1% BSA, 0.05%Tween in PBS (Reagent diluent) was added to each well, and incubated atroom temperature for 1 hr. After 5 times washing, 100 μl ofStreptavidin-conjugated horseradishperoxidase (Genzyme Techne, 1/100 inReagent diluent) was added to each well. After 20 min, each well of theplate was washed 5 times with wash buffer (350 μl/well). The substrateof hourseradishperoxidase and H₂O₂ (TMBZ peroxidase detection kit,SUMILON #ML-1120T) were added to the mixture and the mixture was allowedto stand at room temperature. The reaction was terminated after 10 minby adding 2N H₂SO₄. Optical density at 450 nm was measured with the useof a microplate reader (Labosystems, Multiscan Multisoft).Quantification of TNF-α production in each sample was performed bycomparison of optical densities between each sample and the standardcurve.

The Measurement of IL-2 Production in Jurkat T Cells in Response toAntibody Stimulation

IL-2 production was measured in Jurkat T cells (E6-1 clone; ATCC #TIB-152) in response to stimulation with anti-CD3/anti-CD28 antibodies.

(1) Preparation of Immobilized Antibodies

First, anti-CD3 antibodies (400 ng/well Nichirei, NU-T3 4 μg/ml in 100μl Dulbecco's PBS) were put in each well of 96-well plate (Falcon #3072)and the plate was allowed to stand for 2 hrs at room temperature to becoated with the antibody. Each well of the plate was then washed with250 μl PBS 3 times.

(2) Preparation of Jurkat Cell Culture

Jurkat T cells were cultured in RPMI 1640 medium supplemented with 10%heat-inactivated fetal calf serum, 2 mM L-glutamine, 100 U/ml penicillinG, and 100 μg/ml streptomycin (culture medium). Two hundred thousand(2×10⁵) cells (190 μl/well) were seeded in each well of 96-well U-bottomtissue culture plates (Falcon #3077). To each well was added 10 μlvehicle (0.2% DMSO), serial dilution of compounds in 0.2% DMSO, or 25 nMcyclosporin A as a reference in 0.2% DMSO. The mixture (200 μl) wasincubated for one hour at 37° C. in a humidified 5% CO₂ environment.

(3) Stimulation of the Cell

The reaction mixture obtained in (2) (100 μl) was put in the each wellof the antibody-immobilized plate prepared in (1). To this well wasadded anti-CD28 antibodies (Nichirei, KOLT-2, 6 μg/ml in cell culturemedium, 50 μl/well) and 2.5 μg/ml goat anti-mouse kappa chain antibodies(Bethyl Laboratories, (Cat #A90-119A) 10 μg/ml in culture medium, 50μl/well). The reaction mixture in each well was incubated for 24 hrs at37° C. to stimulate cells with immobilized anti-CD3 antibodies (400ng/well) and anti-CD28 antibodies (1.5 μg/ml), and then to cross-linkreceptors on the cells with anti-mouse kappa chain antibodies (2.5μg/ml).

(4) Measurement of IL-2 Production

The supernatants of the reaction mixture were then collected. The IL-2concentration in the supernatants was determined using a DuoSet™ ELISADevelopment Kit (GenzymeTechne, Minneapolis, USA) following themanufacturer's recommendations. First, 2 μg/ml of mouse anti-huIL-2 Abin PBS buffer (100 μl) was put in each well of 96-well plate (NUNC,Maxisorp™) and the plate was allowed to stand for overnight at 4° C. tobe coated with the antibody. Each well of the plate was then washed 5times with 350 μl of wash buffer containing PBS, 0.05% Tween 20 (Nakalaitesque) using Sera Washer (Bio-Tech, #MW-96R). To each well was added250 μl of 1% BSA (Sigma) in PBS, 0.05% Tween 20 (dilution buffer). After2 hrs incubation at room temperature, the buffer was discarded, and 50μl of culture medium was added. Next, 50 μl supernatant of stimulatedcell culture prepared (3) above was put in each well of the 96-wellplate with coated mouse anti-huIL-2 antibody. Recombinant Human IL-2(Genzyme Techne) was used as the standard for the determination of IL-2production (linear range between 200 and 5,400 μg/ml). The reactionmixtures were incubated for 1 hr at room temperature. After 5 timeswashing, 100 μl biotinylated rabbit anti-huIL-2 antibody (GenzymeTechne, 1.25 μg/ml) in dilution buffer was added to each well, andincubated at room temperature for 1 hr. After 5 times washing, 100 μl ofStreptavidin-conjugated horseradishperoxidase (Genzyme Techne, 1/1000 indilution buffer) was added to each well. After 20 min, each well of theplate was washed 5 times with wash buffer (350 μl/well). Substrate andH₂O₂ (TMBZ peroxidase detection kit, SUMILON #ML-1120T) were added tothe mixture and the mixture was allowed to stand at room temperature.The reaction was terminated after 10 min by adding 2N H₂SO₄. Opticaldensity at 450 nm was measured with the use of a microplate reader(Labosystems, Multiscan Multisoft). Quantification of IL-2 production ineach sample was performed by comparison of optical densities betweeneach sample and the standard curve.

Mouse LPS-induced TNF-α Production

Eight weeks old BALB/c female mice were placed into two groups, acontrol group and a treated group. A solution containing 200 μg/mouse ofLPS in 0.9% physiological salt was administered by intraperitoneal (ip)injection into the control mice. Mice in the treated group were firstinjected ip with compounds of the present invention 30 minutes prior tothe LPS injection. Under anesthesia with pentobarbital (80 mg/kg, i.p.),blood was collected from the posterior venous cavity of the treated andcontrol mice at 90 min post-LPS injection into 96-well plate containing2% EDTA solution. The plasma was separated by centrifugation at 1800 rpmfor 10 minutes at 4° C. and then diluted with four times volumes ofphosphate buffer saline (pH 7.4) containing 1% bovine serum albumin.TNF-α concentration in the sample was determined using an ELISA kit(Pharmingen, San Diego, Calif.)

The mean TNF-α level in 5 mice from each group was determined and thepercent reduction in TNF-α levels was calculated. The treated miceshowed significant decrease in the level of TNF-α as compared to thecontrol mice. The result indicates that the compounds of the presentinvention can restrain LPS-induced cytokine activity.

Results in vitro test and Cellular assay result (A549) are shown inExamples and tables of the Examples below. The data corresponds to thecompounds as yielded by solid phase synthesis and thus to levels ofpurity of about 40 to 90%. For practical reasons, the compounds aregrouped in four classes of activity as follows:

In vitro IC₅₀=A(=or<) 0.5 μM<B (=or<) 2 μM<C (=or<) 10 μM<D

Cellular IC₅₀=A (=or <) 10 μM<B (=or <) 10 μM<C

The compounds of the present invention also show excellent selectivityand strong activity in other cellular activity and in vivo assays.

EXAMPLES

The present invention will be described in detail below in the form ofexamples, but they should by no means be construed as defining the metesand bounds of the present invention.

In the examples below, all quantitative data, if not stated otherwise,relate to percentages by weight. Proton nuclear magnetic resonance (1HNMR) spectra were recorded at either 300 or 500 MHz by Bruker DRX-300

500 Bruker UltraShield™ and chemical shifts are reported in parts permillion relative to tetramethylsilane (TMS). Mass spectra were obtainedusing electrospray (ES) ionization techniques (micromass Platform LC).

[Starting Compound 1A]

A mixture of 2′-hydroxyacetophenone (68.1 g, 0.500 mol), benzylbromide(94.1 g, 0.550 mol) and K₂CO₃ (103 g, 0.750 mol) in acetone (1.0 L) washeated at reflux, and the stirring was continued overnight. After cooledto room temperature, the mixture was concentrated under reducedpressure. The residue was diluted with water, and extracted with ethylacetate. The separated organic phase was washed with brine, dried overMgSO₄, filtered and concentrated under reduced pressure. The crudeproduct was purified by distillation under reduced pressure to give1-[2-(benzyloxy)phenyl]ethanone as a colorless oil. (100 g, yield; 88%)

[Starting Compound 1B]

To a solution of 2′ hydroxyacetophenone (6.79 g, 30 mmol) in ether (200mL) was added a solution of bromine (5.00 g, 31 mmol) in ether (20 mL).After being stirred at room temperature for 30 min, the mixture wasdiluted with ether (100 mL), and then washed with saturated aqueousNaHCO₃ (100 mL) and with brine (100 mL), successively. The combinedorganic phase was dried over Na₂SO₄, filtered and concentrated underreduced pressure. The residue was washed with ether and hexanesuccessively, and dried under reduced pressure to give1-[2-(benzyloxy)phenyl]-2-bromoethanone. (8.42 g, yield; 92%)

To a solution of 1-[2-(benzyloxy)phenyl]-2-bromoethanone (2.59 g, 8.5mmol) and pyridine (1.37 mL, 17 mmol) in THF was addedtetrabutylammonium hydrogen difluoride (4.78 g, 17 mmol). The reactionmixture was stirred under reflux for 22 hrs. The mixture was dilutedwith ether (250 mL), then washed with 1N aqueous hydrochloric acid (100mL) and brine (100 mL), successively. The organic phase was dried overNa₂SO₄, filtered and concentrated under reduced pressure. The crudeproduct was purified by column chromatography on silica gel(hexane/ethyl acetate, 4:1) to give1-[2-(benzyloxy)phenyl]-2-fluoroethanone (688 mg, yield; 33%).

[Starting Compound 1C]

To a stirred solution of 2′-hydroxyacetophenone (5.000 g, 36.724 mmol)in acetonitrile (200 mL) was added potassium carbonate (7.613 g, 55.086mmol). The mixture was stirred at 50° C. for 30 min. 4-Nitrobenzylbromide (8.727 g, 40.396 mmol) was added to the mixture, and thestirring was continued at 50° C. for 12 hrs. Potassium carbonate (0.508g, 3.672 mmol) and 4-nitrobenzyl bromide (0.793 g, 3.672 mmol) wereadded to the mixture and the resulting mixture was stirred at 50° C. for12 hrs. After cooled to room temperature, the reaction mixture wasconcentrated under reduced pressure and diluted with ethyl acetate. Theorganic phase was washed with water, dried over Na₂SO₄, filtered, andconcentrated. The resulting residue was washed with hexane to give1-{2-[(4-nitrobenzyl)oxy]phenyl}ethanoneas a white solid. (8.900 g,yield; 89%)

[Starting Compound 1D]

In a same manner as the method to prepare starting compound 1A, exceptthat benzylbromide was replaced with methoxybenzylchloride,methoxybenzyl-acetophenone was prepared.

[Starting Compound 1E]

A suspension of (2′,6′-dihydroxy)acetophenone (25.0 g, 164 mmol), benzylbromide (40 mL, 337 mmol), potassium carbonate (136 g, 986 mmol) andsodium iodide (2.5 g, 16 mmol) in acetone (500 mL) was stirred at refluxovernight. The mixture was concentrated under reduced pressure, anddiluted with ethyl acetate (500 mL) and water (250 mL). The separatedaqueous phase was extracted with ethyl acetate (200 mL×2). The combinedorganic phase was washed with brine (100 mL), dried over Na₂SO₄,filtered and concentrated under reduced pressure. The residue wastriturated with hexane, collected by filtration, washed with hexane anddried under reduced pressure to give1-[2,6-bis(benzyloxy)phenyl]ethanone (24.9 g, yield; 46%).

[Starting Compound 1F]

A suspension of 3-hydroxypicolinonitrile (3.00 g, 25.0 mmol), which wasprepared according to “Synthesis” 316 (1983) and “J. Org. Chem.” 48 1375(1983), potassium carbonate (5.40 g, 39.1 mmol) and benzyl bromide (5.10g, 29.8 mmol) in acetone (150 mL) was stirred at room temperature for 20hrs. The reaction mixture was filtrated and the filtrate was evaporated.The residue was purified by column chromatography on silica gel (ethylacetate/n-hexane=1/3 to 1/2) followed by recrystallization from ethylacetate/n-hexane=1/4 to give 3-benzyloxypicolinonitrile as a colorlesssolid (4.354 g, yield; 83%).

To a cold (0° C.) solution of 3-benzyloxypicolinonitrile (2.50 g, 11.9mmmol) in tetrahydrofuran (100 mL) was added dropwise 0.92M methylmagnesium bromide in tetrahydrofuran (150 mL, 138 mmol). The mixture wasstirred at 0° C. for 30 minutes and then at room temperature for 4 hrs.The reaction mixture was poured into water (2000 mL) and acidified with10% sulfuric acid (500 mL). After being stirred for 30 min, the reactionmixture was poured into saturate aqueous NaHCO₃ solution slowly andextracted with ethyl acetate. The organic phase was washed with brine,dried over Na₂SO₄, filtrated, and evaporated. The residue was purifiedby column chromatography on silica gel (ethyl acetate/n-hexane=1/3) togive 1-(2-benzyloxy-phenyl)-ethanone as a colorless oil (2.49 g, yield;92%).

[Starting Compound 1G]

To a stirred solution of 1-(2,6-dihydroxyphenyl)ethanone

(50.0 g, 328 mmol) in acetone (1000 mL) was added potassium carbonate(227 g, 1643 mmol) and (bromomethyl)cyclopropane (35.1 mL, 361 mmol).The mixture was stirred at 50° C. for 2 days. The reaction mixture wasfiltrated on Celite®, and then the filtrate was concentrated underreduced pressure. The residue was diluted with water and extracted withethyl acetate. The separated organic phase was washed with water andbrine, dried over MgSO₄, filtered and concentrated under reducedpressure. The residue was suspended in hexane. Then the suspension wasstirred at 80° C. for 30 min. The solution was filtered and the filtratewas allowed to cool to room temperature. The resulting white solid wascollected by filtration, washed with hexane, and dried under reducedpressure to give 1-{2-[(cyclopropylmethyl)oxy]-6-hydroxyphenyl}ethanoneas a pale yellow solid (56.3 g, yield; 83%).

To a stirred solution of1-{2-[(cyclopropylmethyl)oxy]-6-hydroxyphenyl}ethanone (56.3 g, 272mmol) in acetone (1000 mL) was added potassium carbonate (188 g, 1364mmol), 4-methoxybenzyl chloride (40.9 mL, 300 mmol) andtetrabutyl-ammonium iodide (20.2 g, 54.6 mmol). The mixture was stirredat reflux overnight. The reaction mixture was allowed to cool to roomtemperature, filtered on Celite®, and then the filtrate was concentratedunder reduced pressure. The residue was diluted with water and extractedwith ethyl acetate. The separated organic phase was washed with brine,dried over MgSO₄, filtered and concentrated under reduced pressure. Thenthe resulting white solid was recrystallized from ethanol, collected byfiltration, washed with ethanol, and dried under reduced pressure togive 1-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}ethanoneas a white solid (79.2 g, yield; 89%).

[Starting Compound 1H]

To a stirred solution of 5-nitro4H-1,3-benzodioxine (10.0 g, 55.203mmol) in carbon tetrachloride (70 mL) were added N-bromosuccinimide(10.808 g, 60.723 mmol) and 2,2′-azobisisobutyronitrile (0.906 g, 5.520mmol). The mixture was stirred at 100° C. for 2 hrs. After cooled toroom temperature, a solution of sodium ethoxide (4.884 g, 71.764 mmol)in ethanol (70 mL) was added to the mixture, and the stirring wascontinued at room temperature for 3 hrs. The reaction was quenched withwater and extracted with ethyl acetate. The organic phase was washedwith brine, dried over magnesium sulfate, filtered and concentratedunder reduced pressure. The residue was purified by columnchromatography on silica gel (hexane/ethyl acetate=9/1) to give4-ethoxy-5-nitro-4H-1,3-benzodioxine as a pale yellow oil (10.8 g,yield; 87%).

To a stirred solution of 4-ethoxy-5-nitro-4H-1,3-benzodioxine (5.80 g,25.755 mmol) in ethanol (15 mL) and THF (10 mL) was added 4N HCl in 1,4-dioxane (20 mL). The mixture was stirred at 60° C. for 6 hrs and 90°C. for 6 hrs. After cooled to room temperature, the reaction mixture wasdiluted with water and extracted with ethyl acetate. The organic phasewas dried over magnesium sulfate, filtered and concentrated underreduced pressure. The residue was purified by column chromatography onsilica gel (hexane/ethyl acetate=9/1) to give2-hydroxy-6-nitrobenzaldehyde as a pale yellow oil (4.450 g, yield;quant.).

To a stirred solution of 2-hydroxy-6-nitrobenzaldehyde (4.5 g, 26.926mmol) was added dropwise trimethylaluminum (2M in toluene, 27 mL) underan argon atmosphere. The mixture was stirred at room temperature for 1hr. The reaction mixture was poured into a cold (0° C.) aqueous 1Nhydrochloric acid and extracted with ethyl acetate. The organic phasewas washed with brine and dried over Na₂SO₄, filtered and concentrated.The residue was purified by column chromatography on silica gel(hexane/ethyl acetate=9/1) to give 2-(1-hydroxyethyl)-3-nitrophenol asan orange oil (4.660 g, yield; 95%).

To a stirred solution of 2-(1-hydroxyethyl)-3-nitrophenol (0.300 g,1.638 mmol) in acetone (5 mL) was added potassium carbonate (0.25 g,1.802 mmol). The mixture was stirred at room temperature for 15 min, and4-methoxybenzyl chloride (0.22 mL, 1.638 mmol) was added to the mixture.The mixture was stirred at room temperature for 2 hrs and at 60° C. for3 hrs. The reaction was quenched with saturated aqueous ammoniumchloride solution and extracted with ethyl acetate. The organic phasewas dried over Na₂SO₄, filtered and concentrated. The residue waspurified by column chromatography on silica gel (hexane/ethylacetate=4/1). The purified compound was washed with diisopropyl ether togive 1-{2-[(4-methoxybenzyl)oxy]-6-nitrophenyl}ethanol as a white solid(0.201 g, yield; 41%).

To a cooled (0° C.), stirred solution of1-{2-[(4-methoxybenzyl)oxy]-6-nitro-phenyl}ethanol (0.20 g, 0.659 mmol)in dichloromethane (20 mL) were added molecular sieves 4A (1 g),N-methylmorpholine N-oxide (0.15 g, 1.319 mmol) andtetra-n-propylammonium perruthenate (0.01 g). The mixture was stirred atroom temperature for 12 hrs. The reaction mixture was filtered onCelite®, and the filtrate was diluted with ethyl acetate. The organicphase was washed with saturated aqueous ammonium chloride solution anddried over Na₂SO₄, filtered and concentrated under reduced pressure togive 1-{2-[(4-methoxybenzyl)oxy]-6-nitrophenyl}ethanone as a white solid(0.159 g, yield; 80%). The residue was used for the next step withoutfurther purification.

To a stirred suspension of1-{2-[(4-methoxybenzyl)oxy]-6-nitrophenyl}ethanone (0.50 g, 1.659 mmol)in ethanol was added a solution of ammonium chloride (0.10 g, 1.825mmol) in water (5 mL) followed by iron powder (0.75 g). The mixture wasstirred at 100° C. for 1 hr. The reaction mixture was filtrated onCelite®. The filtrate was diluted with ethyl acetate and washed withwater. The separated organic phase was washed with brine and dried overNa₂SO₄, filtrated and concentrated under reduced pressure to give1-{2-amino-6-[(4-methoxybenzyl)oxy]phenyl}ethanone as a yellow solid(0.790 g, yield; 108%). The residue was used for the next step withoutfurther purification.

[Starting Compound 1I]

To a cold (0° C.) solution of 2-fluoro-6-hydroxybenzoic acid (5.00 g,32.029 mmol) in dimethyl formamide (200 mL) were addedN,O-dimethylhydroxylamine hydro-chloride (6.25 g, 64.057 mmol),1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (9.82 g,48.043 mmol), 1-hydroxybenzotriazole (5.19 g, 38.435 mmol), andtriethylamine (8.93 mL, 64.057 mmol). The mixture was stirred at roomtemperature for 15 hrs. The reaction mixture was extracted with ethylacetate and water. The separated organic phase was washed with brine,dried over Na₂SO₄, filtered and concentrated under reduced pressure. Theresulting residue was purified by column chromatography on silica gel(ethyl acetate/hexane=2/1) to give2-fluoro-6-hydroxy-N-methoxy-N-methylbenzamide as an orange solid (4.320g, yield; 68%).

To a stirred solution of 2-fluoro-6-hydroxy-N-methoxy-N-methylbenzamide(4.320 g, 21.689 mmol) in acetone (50 mL) were added 4-methoxybenzylchloride (3.74 g, 23.858 mmol), potassium carbonate (4.50 g,32.534 mmol), and potassium iodide (360 mg, 2.169 mmol). The mixture wasstirred at reflux for 15 hrs. After cooled to room temperature, thereaction mixture was filtered, and the filtrate was concentrated underreduced pressure. The residue was extracted with ethyl acetate andwater. The separated organic phase was washed with brine, dried overNa₂SO₄, filtered and concentrated under reduced pressure. The resultingresidue was purified by column chromatography on silica gel(hexane/ethyl acetate=2/1) to give2-fluoro-N-methoxy-6-[(4-methoxybenzyl)oxy]-N-methylbenzamide as a paleyellow oil (6.926 g, yield; quant.).

To a cold (0° C.) solution of2-fluoro-N-methoxy-6-[(4-methoxybenzyl)oxy]-N-methylbenzamide (6.926 g,21.70 mmol) in tetrahydrofuran (10 mL) was added 1N methyl magnesiumbromide in tetrahydrofuran (43.40 mL, 43.40 mmol). The reaction mixturewas stirred at reflux for 4 hrs. After cooled to room temperature, thereaction mixture was quenched with saturated aqueous NaHCO₃ solution,and extracted with ethyl acetate. The separated organic phase was washedwith water and brine, dried over Na₂SO₄, filtered, and concentratedunder reduced pressure. The resulting residue was purified by columnchromatography on silica gel (hexane:ethyl acetate, 3/1) to give1-{2-fluoro-6-[(4-methoxybenzyl)oxy]phenyl}ethanone as a pale yellowsolid (1.21 g, yield; 20%).

[Starting Compound 1J]

A mixture of methyl 3-hydroxy-2-thiophenecarboxylate (5.00 g, 31.61mmol), benzyl bromide (3.76 mL, 31.61 mmol), and K₂CO₃ (4.81 g, 34.77 g)in acetone (50 mL) was stirred at reflux for 1.5 hrs. After cooled toroom temperature, the mixture was filtered, and the filtrate wasconcentrated under reduced pressure. The residue was purified by columnchromatography on silica gel (hexane:ethyl acetate=15/1 to 9/1) to givemethyl 3-(benzyloxy)-2-thiophenecarboxylate as a pale yellow oil (7.91g, yield: quant.).

To a solution of methyl 3-(benzyloxy)-2-thiophenecarboxylate (7.85 g,31.61 mmol) in MeOH (32 mL) and THF (16 mL) was added 2 N NaOH (21 mL),and the mixture was stirred at reflux for 10 hrs, and then concentratedunder reduced pressure. The residue was diluted with water, washed withether. The separated aqueous phase was acidified with 5N HCl. Theprecipitated solid was collected by filtration, washed with hexane, anddried under reduced pressure at 60° C. to give3-(benzyloxy)-2-thiophenecarboxylic acid as a yellow solid (6.76 g,yield; 91%).

To a solution of 3-(benzyloxy)-2-thiophenecarboxylic acid (3.00 g, 12.81mmol) in CH₂Cl₂ (30 mL) were added oxalyl chloride (1.34 mL, 15.4 mmol)and DMF (0.05 mL). The mixture was stirred at room temperature for 1 hr,and concentrated under reduced pressure. The residue was dissolved inethyl acetate (10 mL). Then the solution was added to a mixture ofN,O-dimethylhydroxyamine hydrochloride (1.50 g, 15.37 mmol) and K₂CO₃(3.54 g, 25.62 mmol) in ethyl acetate (30 mL) and water (30 mL) at 0° C.The mixture was vigorously stirred at 0° C. for 0.5 hrs, and at roomtemperature for 1.5 hrs. The organic phase was separated, washed with 1N HCl, saturated NaHCO₃, and brine, dried over Na₂SO₄, filtered, andevaporated. The residue was purified by column chromatography on silicagel (n-hexane/ethyl acetate=2/1) to give3-(benzyloxy)-N-methoxy-N-methyl-2-thiophenecarboxamide as a pale yellowsolid (3.473 g, yield; 98%).

To a cold (0° C.) solution of3-(benzyloxy)-N-methoxy-N-methyl-2-thiophene-carboxamide (3.47 g, 12.50mmol) in THF (50 mL) was added a solution of methyl-magnesium bromide inTHF (1 M, 35 mL). After being stirred at 0° C. for 1 hr, the reactionmixture was quenched with saturated aqueous NH₄Cl solution, diluted withwater, and extracted with ethyl acetate. The separated organic phase waswashed with brine, dried over Na₂SO₄, filtered and evaporated. Theresidue was purified by column chromatography on silica gel(hexane/ethyl acetate/CH₂Cl₂=3/1/1) followed by recrystallization fromn-hexane/ethyl acetate to give 1-[3-(benzyloxy)-2-thienyl]ethanone as awhite solid (2.592 g, yield; 89%).

[Starting Compound 2A]

To a cold (0° C.) suspension of lithium aluminum hydride (2.44 g, 64mmol) in THF (40 mL) was added methyl1-benzyl-5-oxo-3-pyrrolidinecarboxylate (5.00 g, 21 mmol). The reactionmixture was stirred at room temperature for 5 hrs. To the reactionmixture were added water (2.5 mL), 15% aqueous sodium hydroxide solution(2.5 mL), and water (7.5 mL), successively. The mixture was filteredthrough Celite®. The filtrate was concentrated under reduced pressure togive (1-benzyl-3-pyrrolidinyl)methanol. (4.16 g, yield; quant.)

To a suspension of Pd(OH)₂ (0.4 g) in methanol (30 mL) was added(1-benzyl-3-pyrrolidinyl)methanol (4.16 g, 21 mmol). The reactionmixture was stirred under a hydrogen atmosphere for 24 hours. Thereaction mixture was diluted with ethyl acetate (200 mL), then filteredthrough a Celite®. The filtrate was concentrated under reduced pressureto give 3-pyrrolidinylmethanol (2.33 g, yield; quant.).

To a solution of 3-pyrrolidinylmethanol (2.33 g, 21 mmol) andtriethylamine (4.8 mL, 35 mmol) in dichloromethane (60 mL) was addeddi(tert-butyl) dicarbonate (5.3 g, 24 mmol) at 0° C. The reactionmixture was stirred at room temperature for 26 hrs. The mixture wasdiluted with ethyl acetate (200 mL), and washed with 1N HCl (100 mL),with a saturated aqueous NaHCO₃ solution (100 mL) and with brine (100mL), successively. The organic phase was dried over Na₂SO₄, filtered,and concentrated under reduced pressure to give tert-butyl3-(hydroxymethyl)-1-pyrrolidinecarboxylate (4.06 g, yield; 96%).

To a cold (0° C.) mixture of tert-butyl3-(hydroxymethyl)-1-pyrrolidinecarboxylate (4.0 g, 20 mmol),dichloromethane (100 mL), dimethyl sulfoxide (20 mL) and tri-ethylamine(16.9 mL, 121 mmol) under an argon atmosphere was added sulfurtrioxide-pyridine complex (9.63 g, 60 mmol). The reaction mixture wasallowed to warm to room temperature, and the stirring was continued for1 hr. The mixture was extracted with ether (200 mL) and a saturatedaqueous NaHCO₃ solution (100 mL). The separated aqueous phase wasfurther extracted with ether (100 mL×2). The combined organic phase waswashed with a 1N aqueous HCl solution (100 mL), a saturated aqueousNaHCO₃ solution (100 mL), and brine (100 mL), successively. The organicphase was dried over Na₂SO₄, filtered and concentrated under reducedpressure (5.32 g, yield; quant.).

[Starting Compound 2B]

To a cooled (0° C.), stirred solution of 3-piperidinecarboxylic acid(100.000 g, 774.233 mmol) in dioxane (400 mL) were added 2N NaOH (400mL, 800 mmol) and di-tert-butyl dicarbonate (168.978 g, 774.233 mmol).The mixture was allowed to warm to room temperature, and the stirringwas continued for 12 hrs. The mixture was concentrated under reducedpressure. The residue was diluted with water and acidified (pH 3-4) withaqueous 1N HCl solution. The resulting solid was collected byfiltration. The white solid was dissolved in ethyl acetate and washedwith water. The separated organic phase was dried over Na₂SO₄, filtered,and concentrated. The resulting solid was suspended in hexane andcollected by filtration, and dried under reduced pressure to give1-(tert-butoxycarbonyl)-3-piperidine carboxylic acid as a white solid.(156 g, yield; 88%)

To a cold (0° C.) solution of 1-(tert-butoxycarbonyl)-3-piperidinecarboxylic acid (7.000 g, 30.531 mmol) in dichloromethane (200 mL)including triethylamine (4.681 mL, 33.584 mmol) were addedbenzotriazole-1-yl-oxy-tris-pyrrolidino-phosphonium hexafluorophosphate(15.885 g, 30.531 mmol), N,O-dimethylhydroxyamine (3.276 g, 33.584 mmol)and triethylamine (4.255 mL, 30.531 mmol) successively. The mixture wasallowed to warm to room temperature, and the stirring was continued for12 hrs. The reaction mixture was diluted with dichloromethane and washedwith an aqueous 1N HCl solution, saturated aqueous NaHCO₃ solution, andbrine, successively. The organic phase was dried over Na₂SO₄, filtered,and concentrated. The resulting residue was purified by columnchromatography on silica gel (chloroform/ethyl acetate=10/1-9/1) to give3-{[methoxy(methyl)amino]carbonyl}-1-piperidine-carboxylic acidtert-butyl ester as a white solid. (8.050 g, yield; 96%)

To a cooled (−15° C.), stirred suspension of lithium aluminum hydride(4.355 g, 114.743 mmol) in diethyl ether (500 mL) was added dropwise asolution of 3-{[methoxy(methyl)amino]carbonyl}-1-piperidine-carboxylicacid tert-butyl ester (25.000 g, 91.795 mmol) in THF (150 mL) over 30min. The reaction mixture was quenched with aqueous 1N potassiumhydrogen sulfate (300 mL), and extracted with a 1:1 mixture of diethylether and ethyl acetate. The organic phase was dried over MgSO₄,filtered, and concentrated under reduced pressure to give3-formyl-piperidine-1-carboxylic acid tert-butyl ester, which was usedfor the next steps without further purification. (22.56 g, yield;quant.)

[Starting Compound 2C]

To a cooled (0° C.) and stirred solution of nipecotinic acid (3.0 g,23.3 mmol) in 1,4-dioxane (12 mL) was added 2N NaOH solution (24.0 mL,48.0 mmol) followed by a solution of benzyl chloroformate (3.96 g, 23.2mmol) in 1,4-dioxane (12 mL). The reaction mixture was allowed to warmto room temperature, and the stirring was continued for 3 hrs. Themixture was concentrated under reduced pressure. The residue was dilutedwith water and acidified with 1H HCl (pH 3-4). The mixture was extractedwith ethyl acetate, and the separated organic phase was washed withbrine, dried over Na₂SO₄, filtered, and evaporated. The resulting whitesolid was suspended in hexane, collected by filtration, washed withhexane, and dried under reduced pressure to give1-[(benzyloxy)carbonyl]-3-piperidinecarboxylic acid as a white solid(4.4 g, yield; 71%).

To a cooled (−15° C. to −20° C.) and stirred solution of1-[(benzyloxy)carbonyl]-3-piperidinecarboxylic acid (4.0 g, 15.2 mmol)in dry THF (50 mL) including methylmorpholine (2 mL) was added dropwisea solution of isobutyl chioroformate (2.28 g, 16.7 mmol) in THF (10 mL).After the mixture was stirred at the same temperature for 20 min, asolution of N,O-dimethylhydroxylamine hydrochloride (1.63 g, 16.7 mmol)in THF (20 mL) including methylmorpholine (2 mL) was added. The reactionmixture was allowed to warm to room temperature, and then stirred for 16hrs. The reaction mixture was concentrated under reduced pressure andpartitioned between ethyl acetate and water. The separated organic phasewas washed with 1N HCl, saturated NaHCO₃, and brine, successively. Theorganic phase was dried over Na₂SO₄, filtered, and concentrated underreduced pressure to give benzyl3-{[methoxy(methyl)amino]carbonyl)-1-piperidinecarboxylate as acolorless oil (4.7 g, yield; quant.).

To a cooled (−78° C.) suspension of lithium aluminum hydride (0.70 g,18.4 mmol) in dry THF (100 mL) under an argon atmosphere was addeddropwise a solution of benzyl3-{[methoxy(methyl)amino]carbonyl}-1-piperidinecarboxylate (4.5 g, 14.7mmol) in THF (30 mL), and the stirring was continued at −78° C. for 45min. The reaction mixture was quenched with 0.5N KHSO₄ (130 mL), andthen extracted with ether. The separated organic phase was dried overMgSO₄, filtered, and concentrated under reduced pressure to give benzyl3-formyl-1-piperidinecarboxylate as a colorless oil (0.54 g, yield;82%).

To a cold (0° C.) mixture of 3-aminocyclohexane-carboxylic acid (10.0 g,69.8 mmol) in 1,4-dioxane (100 mL) and 2N NaOH (100 mL) was addeddi(tert-butyl) dicarbonate (15.2 g, 69.8 mmol). The mixture was stirredat room temperature for 1.5 hrs, and then neutralized with 2N HCl. Theresulting mixture was concentrated under reduced pressure. The residuewas acidified with 2N HCl and extracted with ethyl acetate. Theseparated organic phase was washed with brine, dried over Na₂SO₄,filtered and evaporated. The residual solid was triturated with hexane,collected by filtration, and dried under reduced pressure to give3-[(tert-butoxycarbonyl)amino]cyclohexanecarboxylic acid as a whitesolid (8.00 g, yield; 47%).

To a cold (0° C.) solution of3-[(tert-butoxycarbonyl)amino]-cyclohexanecarboxylic acid (5.00 g, 20.6mmol) in CH₂Cl₂ (50 mL) including triethylamine (3.1 mL, 22.2 mmol) wereadded benzotriazol-1-yloxytris(pyrrolidino)phosphoniumhexa-fluorophosphate (10.7 g, 20.6 mmol), N,O-dimethylhydroxyaminehydrochloride (2.21 g, 20.6 mmol) and Et₃N (3.2 mL, 23.0 mmol)successively. The resulting mixture was allowed to warm to roomtemperature, and the stirring was continued for 18 hrs. The reactionmixture was diluted with CH₂Cl₂ and washed with 1N HCl, saturatedaqueous NaHCO₃ solution, and brine, successively. The organic phase wasdried over Na₂SO₄, filtered, and evaporated. The residue was purified bycolumn chromatography on silica gel (hexane/ethyl acetate=1:1) to givetert-butyl 3-([methoxy(methyl)amino]carbonyl}cyclohexylcarbamate as acolorless oil (5.39 g, yield; 92%).

To a suspension of NaH (60%, 340 mg, 8.38 mmol) in THF (10 mL) was addeddropwise a solution of tert-butyl3-{[methoxy(methyl)amino]carbonyl}cyclohexylcarbamate (2.00 g, 6.98mmol) in THF (10 mL)followed by methyl iodide (1.09 g, 7.68 mmol). Themixture was stirred at 60° C. for 0.5 hr. After cooled to roomtemperature, the mixture was poured into water and extracted with ethylacetate. The organic phase was washed with brine, dried over Na₂SO₄,filtered and evaporated. The residue was purified by columnchromatography on silica gel (hexane/ethyl acetate=1/1) to givetert-butylN-[3-(N-methoxy-N-methyl-carbamoyl)cyclohexyl]-N-methylcarbamate as acolorless oil (1.94 g, yield; 93%).

To a cold (0° C.) suspension of LiAlH₄ (380 mg, 9.9 mmol) in Et₂O (40mL) was added dropwise a solution of tert-butylN-[3-(N-methoxy-N-methylcarbamoyl)cyclohexyl]-N-methylcarbamate (2.38 g,7.92 mmol) in Et₂O (40 mL) over 10 min. The reaction mixture was stirredat 0° C. for 0.5 hr, and quenched with 1N KHSO₄ (100 mL). The organicphase was separated, dried over MgSO₄ and evaporated to give tert-butyl3-formylcyclohexyl(methyl)carbamate as a colorless oil, which was usedfor next step without purification (1.66 g, yield; 87%).

[Starting Compound 2E]

To a cold (0° C.) solution of ethyl nipecotate (7.86 g, 50 mmol) inCH₂Cl₂ (120 mL) was added di-tert-butyl dicarbonate (11.46 g, 52.5mmol). The resulting mixture was stirred at room temperature for 4 hrs.The reaction mixture was diluted with a 5% aqueous NaHCO₃ solution. Theorganic phase was separated, dried over MgSO₄, filtered and evaporatedunder reduced pressure to give 1-tert-butyl 3-ethyl1,3-piperidinedicarboxylate (12.8 g, yield; quant.).

To a cold (−78° C.) solution of lithium diisopropylamide (LDA) (23.3mmol) in THF (10 mL) under an argon atmosphere was added dropwise asolution of 1-tert-butyl 3-ethyl 1,3-piperidinedicarboxylate (3.0 g,11.6 mmol) in THF (5 mL), then stirred at −50° C. for 2 hrs. To theresulting mixture was added a solution of MeI (1.98 g, 14 mmol) in THFat −50° C. The reaction mixture was allowed to warm to room temperature,and quenched with saturated aqueous NH₄Cl solution. The mixture wasconcentrated under reduced pressure, and the residue was partitionedbetween ethyl acetate and water. The organic phase was separated, driedover MgSO₄, filtered and concentrated under reduced pressure. The crudeproduct was purified by column chromatography on silica gel(hexane:ethyl acetate, 9:1) to give 1-tert-butyl 3-ethyl3-methyl-1,3-piperidinedicarboxylate as a pale yellow oil (2.27 g,yield; 72%).

To a cold (−20° C.) solution of 1-tert-butyl 3-ethyl3-methyl-1,3-piperidinedi-carboxylate (2.3 g, 8.48 mmol) in dry THF wasadded LiBH₄ (2M in THF, 10.6 mmol), and the stirring was continuedovernight. The reaction mixture was allowed to warm to room temperature.The reaction mixture was cooled by ice water and acidified (pH 5-6) withaqueous 1N HCl. The mixture was concentrated under reduced pressure,then extracted with ethyl acetate. The organic phase was separated,dried over MgSO₄, filtered, and evaporated. The crude product waspurified by column chromatography on silica gel (hexane:ethyl acetate,4:1-3:1) to give tert-butyl3-(hydroxymethyl)-3-methyl-1-piperidinecarboxylate as a colorless oilform (1.5 g, yield; 78%).

To an ice water cooled solution of tert-butyl3-(hydroxymethyl)-3-methyl-1-piperidinecarboxylate (1.5 g, 6.6 mmol) inCH₂Cl₂ were added DMSO (7.1 g, 91.6 mmol), Et₃N (5.5 mL, 39.8 mmol), andsulfur trioxide-Pyridine complex (3.1 g, 19.9 mmol) successively. Themixture was stirred at room temperature for 1 hr. The reaction mixturewas diluted with ether, purified by column chromatography on silica gel(hexane:ethyl acetate, 5:1-4:1) to give tert-butyl3-formyl-3-methyl-1-piperidinecarboxylate as a colorless oil form (1.3g, yield; 86%).

To a solution of 1,3-cyclohexanedicarboxylic acid (10 g, 58 mmol) inMeOH (50 mL) was added conc. H₂SO₄ (2 mL). Then the mixture was stirredat room temperature for 4 hrs. The mixture was concentrated underreduced pressure, and then partitioned between ethyl acetate and asaturated aqueous NaHCO₃ solution. The organic phase was washed withbrine, dried over Na₂SO₄, and evaporated to give dimethyl1,3-cyclohexanedicarboxylate as a colorless oil (11.6 g, yield; quant.).To a solution of dimethyl 1,3-cyclohexanedicarboxylate (11.6 g, 57.9mmol) in MeOH (58 mL) was added a 1N NaOH solution (58 mL) dropwise over1 hr at 0° C. The resulting mixture was stirred at 0° C. for 0.5 hrs,and at room temperature for 2 hrs. The mixture was concentrated underreduced pressure, and the residual solution was partitioned betweenethyl acetate and water. The aqueous phase was separated, acidified withconc. HCl (15 mL), saturated with NaCl, and then extracted with ethylacetate. The extract was dried over Na₂SO₄, filtered and concentratedunder reduced pressure to give 3-(methoxycarbonyl)cyclohexanecarboxylicacid as a colorless oil (6.16 g, yield; 57%).

To a cold (−78° C.) solution of 3-(methoxycarbonyl)cyclohexanecarboxylicacid (1.25 g, 6.71 mmol) in THF (20 mL) was added BH₃.Me₂S (0.7 mL, 7.38mmol). The resulting mixture was stirred at −78° C. to room temperatureovernight. The reaction mixture was quenched with a saturated aqueousNH₄Cl solution and extracted with ethyl acetate. The organic phase waswashed with brine, dried over Na₂SO₄, and evaporated. The residue waspurified by column chromatography on silica gel (n-hexane/ethylacetate=2/1) to give methyl 3-(hydroxymethyl)cyclo-hexanecarboxylate asa colorless oil (845 mg, yield; 73%). To a solution of methyl3-(hydroxymethyl)cyclohexanecarboxylate (840 mg, 4.88 mmol) in Et₃N (4.4mL, 32 mmol) and DMSO (10 mL) was added sulfur trioxide-pyridine complex(2.56 g, 16.1 mmol). The mixture was stirred at room temperature for 1hr. The reaction mixture was partitioned between ethyl acetate andwater. The organic phase was washed with brine, dried over Na₂SO₄,filtered and evaporated. The residue was purified by columnchromatography on silica gel (n-hexane/ethyl acetate=2/1) to give methyl3-formylcyclohexanecarboxylate as a pale yellow oil (795 mg, yield;96%).

To a suspension of methoxymethylphosphonium chloride (5.14 g, 15 mmol)in THF (18 mL) was added potassium tert-butoxide (1.98 g, 15 mmol) at 0°C., and the mixture was stirred at 0° C. for 0.5 hr. A solution oftert-butyl 3-formyl-1-piperidinecarboxylate (2.13 g, 10 mmol) in THF (8mL) was added dropwise at −20° C., and the resulting mixture was stirredat 0° C. to room temperature overnight. The mixture was partitionedbetween toluene and water. The separated organic phase was washed withbrine, dried over Na₂SO₄, filtered, and evaporated. Purification bycolumn chromatography on silica gel (hexane/ethyl acetate=5/1) gavetert-butyl 3-(2-methoxyethenyl)-1-piperidinecarboxylate as a colorlessoil (1.16 g, yield; 48%).

tert-Butyl 3-(2-methoxyethenyl)-1-piperidinecarboxylate (1.16 g, 4.81mmol) was dissolved in 90% formic acid (2 mL). The mixture was stirredat room temperature for 40 min. The mixture was then partitioned betweenethyl acetate and saturated aqueous NaHCO₃ solution. The separatedorganic phase was washed with brine, dried over Na₂SO₄, filtered andevaporated. Purification by column chromatography (hexane/ethylacetate=4/1) gave tert-butyl 3-(2-oxoethyl)-1-piperidinecarboxylate as acolorless oil (652 my, yield; 60%).

To a cold (0° C.) mixture of 3-methoxycarbonyl 4-piperidone HCl (10 g,51.6 mmol) and triethylamine (10.8 mL, 77.5 mmol) in dichloromethane(100 mL) was added a solution of di-tert-butyl dicarbonate (11.8 g, 54.2mmol) in dichloromethane (100 mL). The mixture was stirred at roomtemperature for 2 hrs and concentrated under reduced pressure. Theresidue was quenched with water, extracted with ether, dried over MgSO₄,filtered, and evaporated to give 1-tert-butyl 3-methyl4-oxo-1,3-piperidinedicarboxylate, which was used for the next reactionwithout further purification. (14.9 g, yield; quant.)

To a cold (0° C.) solution of 1-tert-butyl 3-methyl4-oxo-1,3-piperidinedicarboxylate (134.0 g, 520.8 mmol) in methanol (800mL) was added NaBH₄ (9.85 g, 260.4 mmol) portionwise. The resultingmixture was stirred overnight to be allowed to warm to room temperature.The solvent was removed under reduced pressure, the residue wasdissolved into ethyl acetate, washed with brine, dried over MgSO₄,filterd, and concentrated under reduced pressure to give 1-tert-butyl3-methyl 4-hydroxy-1,3-piperidinedicarboxylate, which was used for thenext reaction without further purification. (120.8 g, yield 90%)

To a cold (0° C.) solution of 1-tert-butyl 3-methyl4-hydroxy-1,3-piperidine-dicarboxylate (1,64 g, 6.32 mmol),4-dimethylaminopyridine (16.0 mg, 0.13 mmol), and triethylamine (2.64mL, 19.0 mmol) in dichloromethane (25 mL) was added dropwisetrifluoroacetic anhydride (1.12 mL, 7.91 mmol). The reaction mixture wasstirred at room temperature for 40 hrs. The reaction was quenched with10% K₂CO₃ solurtion, extracted with CHCl₃. The separated organic phasewas washed with brine, dried over MgSO₄, filtered and evaporated underreduced pressure. The crude product was purified by silica gel columnchromatography (hexane:ethyl acetate=5:1) to give 1-tert-butyl 3-methyl5,6-dihydro-1,3(2H)-pyridinedicarboxylate as a pale yellow oil. (707.4mg, yield 46%)

To a cold (−20° C.) solution of 1-tert-butyl 3-methyl5,6-dihydro-1,3(2H)-pyridinedicarboxylate (700.0 mg, 2.90 mmol) in drytoluene (40 mL) was added a solution of diisobutylaluminum hydride intoluene (1.5 M, 4.83 mL, 7.25 mmol) dropwise under an argon atmosphere.The reaction mixture was stirred at 0° C. for 30 min and then stirred atroom temperature. The reaction was quenched by an addition of water andthe stirring was continued for additional 1 hr. The precipitatedaluminum salts were filtered off. The filtrate was diluted withsaturated NH₄Cl solution, extracted with ethyl acetate. The separatedorganic phase was washed with brine, dried over MgSO₄, filtered andevaporated to give tert-butyl5-(hydroxymethyl)-3,6-dihydro-1(2H)-pyridinecarboxylate, which was usedfor the next reaction without further purification. (580.0 mg, yield94%)

To a solution of tert-butyl5-(hydroxymethyl)-3,6-dihydro-1(2H)-pyridinecarboxylate (14.65 g, 68.69mmol) in dichloromethane was added MnO₂(89.58 g, 1030.36 mmol), thenstirred overnight at room temperature. The reaction mixture was filteredthrough Celite®, the filtrate was concentrated under reduced pressure togive tert-butyl 5-formyl-3,6-dihydro-1(2H)-pyridinecarboxylate, whichwas pure enough to be used for the next reaction. (13.55 g, yield 94%)

Example 1-1

(1) A mixture of 2′-benzyloxyacetophenone (starting compound 1A) (10.00g, 44.19 mmol), tert-butyl 3-formyl-1-piperidine carboxylate (startingcompound 2B) (10.37 g, 44.61 mmol), malononitrile (3.21 g, 48.61 mmol),and ammonium acetate (17.03 g, 220.97 mmol) in toluene (50 mL) wasstirred at 150° C. for 2 hrs. After cooled to room temperature, thereaction mixture was diluted with ethyl acetate. The organic phase waswashed with water and saturated brine, dried over Na₂SO₄, filtered andconcentrated. The residue was purified by column chromatography onsilica gel (hexane/ethyl acetate=2/1). The purified compound (yellowoil) was recrystallized from ethanol to give tert-butyl 3-{2-amino-6-[2-(benzyloxy)phenyl]-3-cyano-4-pyridinyl}-1-piperidinecarboxylateas a white solid. (5.83 g, yield; 27%).

(2) A mixture of tert-butyl3-{2-amino-6-[2-(benzyloxy)phenyl]-3-cyano-4-pyridinyl}-1-piperidinecarboxylate(2.160 g, 4.457 mmol) and 10% Pd—C (0.720 g) in ethyl acetate (30 mL)was stirred at room temperature for 2 days under a hydrogen atmosphere(3 atm). The mixture was filtrated with Celite® and the filtrate wasconcentrated under reduced pressure. The residue was recrystallized fromdiethyl ether to give tert-butyl3-[2-amino-3-cyano-6-(2-hydroxyphenyl)-4-pyridinyl]-1-piperidinecarboxylateas a white solid. (0.180 g, yield; 10%)

(3) To a stirred solution of tert-butyl3-[2-amino-3-cyano-6-(2-hydroxyphenyl)-4-pyridinyl]-1-piperidinecarboxylate(0.170 g, 0.431 mmol) in dioxane (30 mL) was added 4N HCl in dioxane (3mL). The mixture was stirred at room temperature for 12 hrs. Theresulting precipitates were collected by filtration and the filtrate waswashed with dioxane to give2-amino-6-(2-hydroxyphenyl)-4-(3-piperidinyl)-nicotinonitrilehydrochloride (0.075 g, yield; 53%).

Molecular weight: 330.82; Mass spectrometry: 295 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A/(Jurkat)-A;¹H-NMR(300 MHz, DMSO-d6): 1.81-1.89 (4H, m), 2.88-2.92 (1H, m),3.31-3.40 (4H, m), 6.89-6.94 (2H, m), 7.32-7.38 (1H, m), 7.43 (1H, s),8.05 (1H, d, J=7.4 Hz), 8.83-8.85 (1H, br s), 9.45-9.47 (1H, br s).

Example 1-2

With the use of starting compound 2A instead of 2B,2-amino-6-(2-hydroxyphenyl)-4-(3-pirrolidinyl)nicotinonitriletrifluoroacetate was prepared in a similar manner as described inExample 1-1.

Molecular weight: 394.36; Mass spectrometry: 281 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A594)-A/(Jurkat)-B;¹H-NMR(500 MHz, DMSO-d6): 2.10 (1H, m), 3.60-3.70 (2H, m), 6.91 (2H, m),7.36-7.44 (4H, m), 8.05 (1H, d, J=8.2 Hz), 9.00 (2H, br), 13.35 (1H, s).

Example 1-3

With the use of starting compound 2E instead of 2B,2-amino-6-(2-hydroxyphenyl)-4-(3-methylpiperidin-3-yl)-nicotinonitrilehydrochloride was prepared in a similar manner as described in Example1-1.

Molecular weight: 344.85; Mass spectrometry: 309 (M+H)⁺; In vitroactivity grade: B; Cellular activity grade: (A594)-B; ¹H-NMR(500 MHz,DMSO-d6): 1.52 (3H, s), 1.78 (2H, br d, J=38.1 Hz), 1.97-2.04 (1H, m),2.37-2.44 (1H, m), 2.99 (2H, br d, J=31.7 Hz), 3.34 (1H, dd, J=5.5, 12.5Hz), 3.75 (1H, dd, J=5.5, 12.5 Hz), 6.92-6.96 (2H, m), 7.22 (1H, s),7.33-7.39 (2H, m), 8.05-8.08 (1H, m), 9.02 (1H, br s), 9.42 (1H, br s).

Example 1-4

With the use of the starting compound 1B instead of 1A,2-amino-5-fluoro-6-(2-hydroxyphenyl)-4-(3-piperidinyl)-nicotinonitriletrifluoroacetate was prepared in a similar manner as described inExample 1-1.

Molecular weight: 426.37; Mass spectrometry: 313 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A594)-B/(Jurkat)-B;¹H-NMR(500 MHz, DMSO-d6): 1.77 (2H, m), 1.95 (3H, m), 2.94 (1H, t,J=10.7 Hz), 3.25-3.37 (2H, m), 3.50 (1H, d, J=12.0 Hz), 6.90 (1H, dt,J=1.0, 8.2 Hz), 6.93 (1H, d, J=8.2 Hz), 7.03 (2H, br), 7.32 (1H, dt,J=1.6, 8.5 Hz), 7.47 (1H, dd, J=1.6, 7.9 Hz), 8.73 (2H, br), 10.72 (1H,s).

Example 1-5

With the use of the starting compound 2D instead of 2B,2-amino-6-(2-hydroxyphenyl)-4-[3-methylamino]cyclohexyl]nicotinonitrilehydrochloride was prepared in a same manner as described in Example 1-1.

Molecular weight: 358.87; Mass spectrometry: 323 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A594)-B; ¹H-NMR(300 MHz,DMSO-d6): 1.34-1.86 (5H, m), 1.86-2.04 (1H, m), 2.04-2.30 (2H, m),2.50-2.60 (3H, m), 2.76-2.95 (1H, m), 3.17 (1H, br s), 6.85-6.96 (2H,m), 7.20-7.40 (3H, m), 7.99 (1H, d, J=8.1 Hz), 9.11 (2H, br s).

Example 1-6

With the use of starting compound 1F instead of 1A,6-amino-3′-hydroxy-4-(3-piperidinyl)-2,2′-bipyridine-5-carbonitrilehydrochloride was prepared in a same manner as described in Example 1-1.

Molecular weight: 368.27; Mass spectrometry: 337 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A594)-A; ¹H-NMR (500 MHz,DMSO-d6): 1.78-1.82 (2H, m), 1.93-1.95 (2H, m), 3.03-3.07 (1H, m),3.27-3.37 (3H, m), 3.56 (1H, m), 7.39 (1H, dd, J=1.6, 8.5 Hz), 7.43 (1H,dd, J=4.1, 8.5 Hz), 7.56 (1H, br), 7.76 (1H, s), 8.23 (1H, dd, J=1.6,4.1 Hz), 8.78 (1H, br), 9.18 (1H, br).

Example 1-7

With the use of the starting compound 1J instead of 1A,2-amino-6-(3-hydroxy-2-thienyl)4-(3-piperidinyl)nicotinonitrilehydrobromide was prepared in a same manner as described in Example 1-1.

Molecular weight: 381.30; Mass spectrometry: 301 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A594)-A; ¹H-NMR (500 MHz,DMSO-d6): 1.69-1.84 (2H, m), 1.84-2.00 (2H, m), 2.94-3.06 (1H, m),3.12-3.28 (2H, m), 3.28-3.40 (2H, m), 6.82 (2H, d, J=5.4 Hz), 6.83 (1H,s), 7.63 (2H, d, J=5.4 Hz), 8.55-8.70 (1H, m), 8.85-9.00 (1H, m).

Example 1-8

With the use of the starting compound 2F instead of 2B, methyl3-[2-amino-3-cyano-6-(2-hydroxyphenyl)4-pyridinyl]-cyclohexanecarboxylatewas prepared in a similar manner as described in Example 1-1.

A suspension of methyl3-[2-amino-3-cyano-6-(2-hydroxyphenyl)-4-pyridinyl]cyclohexanecarboxylate(151 mg, 0.43 mmol) in 2N NaOH (1 mL) and THF (2 mL) was stirred at 60°C. for 4 hrs. The reaction mixture was neutralized with 1N HCl (2 mL),and then diluted with water. The resulting precipitates were collectedby filtration, washed with EtOH, and dried (60° C., 2 hrs, under reducedpressure) to give3-[2-amino-3-cyano-6-(2-hydroxyphenyl)-4-pyridinyl]cyclohexanecarboxylicacid as a yellow solid (105 mg, yield; 72%).

Molecular weight: 337.38; Mass spectrometry: 338 (M+H)⁺; In vitroactivity grade: B; Cellular activity grade: (A594)-C; ¹H-NMR (300 MHz,DMSO-d6): 1.30-2.14 (8H, m), 2.30-2.59 (1H, m), 2.70-2.89 (1H, m),6.83-6.93 (2H, m), 7.24-7.38 (4H, m), 8.00-8.10 (1H, m), 12.41 (1H, s),13.47 (1H, s).

Examples 1-9

With the use of the starting compound 1D instead of 1A and the startingcompound 2H instead of 2B,2′-amino-6′-(2-hydroxyphenyl)-1,2,5,6-tetrahydro-3,4′-bipyridine-3′-carbonitrilehydrochloride was prepared in a same manner as described in Example 1-1.

Molecular weight: 328.80; Mass spectrometry: 293 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A594)-A; ¹H-NMR (500 MHz,DMSO-d6): 2.52 (2H, br s), 3.24 (2H, br s), 4.05(2H, br s), 6.45 (1H,s), 6.90-6.93 (2H, m), 7.35-7.46 (4H, m), 8.01-8.02 (1H, m), 9.37 (1H,br s).

Examples 1-10

With the use of starting compound 1H instead of 1A,2-amino-6-(2-amino-6-hydroxyphenyl)4-(3-piperidinyl)nicotinonitrilehydrochloride was prepared in a same manner as described in Example 1-1.

Molecular weight: 345.83; Mass spectrometry: 310 (M+H)⁺; In vitroactivity grade: C; Cellular activity grade: (A594)-C; ¹H-NMR (500 MHz,DMSO-d6): 1.81-1.92 (3H, m), 2.89-2.93 (1H, m), 3.28-3.32 (3H, m), 6.97(2H, dd, J=11.7, 8.2 Hz), 7.30 (2H, t, J=8.2 Hz), 9.01 (1H, m), 9.59(1H, m).

Examples 1-11

(1) With the use of the starting compound 1H instead of 1A, tert-butyl3-(2-amino-6-{2-amino-6-[(4-methoxybenzyl)oxy]phenyl}-3-cyano-4-pyridinyl)-1-piperidinecarboxylatewas prepared in a same manner as described in the step (1) of Example1-1.

(2) To a cooled (0° C.), stirred solution of tert-butyl3-(2-amino-6-{2-amino-6-[(4-methoxybenzyl)oxy]phenyl}-3-cyano-4-pyridinyl)-1-piperidinecarboxylate(0.30 g, 0.566 mmol) in methanol (10 mL) including acetic acid (0.03 g,0.566 mmol) was added benzaldehyde (0.08 mL, 1.133 mmol) followed bysodium cyanoborohydride (0.04 g, 0.566 mmol), and the stirring wascontinued at 0° C. for 3 hrs. The reaction was quenched with water andextracted with ethyl acetate. The organic phase was dried over sodiumsulfate, filtered and concentrated under reduced pressure. The residuewas purified by silica gel column chromatography (hexane/ethylacetate=4/1) to give tert-butyl3-(2-amino-6-{2-(benzylamino)-6-[(4-methoxybenzyl)oxy]-phenyl}-3-cyano-4-pyridinyl)-1-piperidinecarboxylateas a yellow amorphous (0.299 g, yield; 85%).

(3) Then the yellow amorphous was treated under acidic conditions in thesimilar manner as that of the step (3) in Example 1-1 to give2-amino-6-[2-(benzylamino)-6-hydroxyphenyl]-4-(3-piperidinyl)nicotinonitrilehydrochloride.

Molecular weight: 435.96; Mass spectrometry: 400 (M+H)⁺; In vitroactivity grade: B; Cellular activity grade: (A594) C; ¹H-NMR (500 MHz,DMSO-d6): 1.69-1.83 (2H, m), 1.90-1.92 (2H, m), 2.89-2.92 (1H, m),3.15-3.18 (1H, m), 3.28-3.33 (3H. m), 4.28 (2H, s), 6.17 (1H, d, J=8.2Hz), 6.32 (1H, d, J=8.2 Hz), 7.00 (1H, t, J=8.2 Hz), 7.09 (1H, s),7.20-7.23 (1H, m), 7.28-7.33 (4H, m), 8.97 (1H, m), 9.47 (1H, m).

Example 1-12

To a cooled (0° C.), stirred solution of tert-butyl3-(2-amino-6-{2-amino-6-[(4-methoxybenzyl)oxy]phenyl}-3-cyano-4-pyridinyl)-1-piperidinecarboxylate(0.10 g, 0.189 mmol) obtained in the step (1) of Example 1-11 indichloromethane (10 mL) including triethylamine (0.02 g, 0.227 mmol) wasadded acetic anhydride (0.08 g, 0.889 mmol).The stirring was continuedat 0° C. for 5 hrs. The mixture was allowed to warm to room temperature,and the stirring was continued for 30 min. The reaction was quenchedwith water and extracted with ethyl acetate. The organic phase waswashed with brine and dried over Na₂SO₄, filtered and concentrated underreduced pressure. The residue was purified by silica gel columnchromatography (hexane/ethyl acetate=2/1-1/1-1/2) to give tert-butyl3-(6-{2-(acetylamino)-6-[(4-methoxybenzyl)oxy]phenyl}-2-amino-3-cyano-4-pyridinyl)-1-piperidine-carboxylateas a pale yellow oil (0.102 g, yield; 94.5%).

Then the yellow oil was treated with acids in the same manner as in thestep (2) in Example 1-1 to giveN-{2-[6-amino-5-cyano-4-(3-piperidinyl)-2-pyridinyl]-3-hydroxyphenyl}acetamidehydrochloride.

Molecular weight: 387.87; Mass spectrometry: 352 (M+H)⁺; In vitroactivity grade: C; Cellular activity grade: (A594) C; ¹H-NMR (500 MHz,DMSO-d6): 1.60-1.86 (4H, m), 1.90 (3H, s), 2.88-2.95 (1H, m), 3.12-3.26(4H, m), 6.78 (1H, d, J==8.2 Hz), 7.02 (1H, s), 7.23 (1H, t, J=8.2 Hz),7.28-7.30 (1H, m), 8.90-8.92 (1H, m), 9.40-9.42 (1H, m), 9.90 (1H, brs).

Examples 1-13 to 1-44

In similar manners as described in Example 1-1 to Example 1-12 above,compounds in Examples 1-13 to 1-44 as shown in Table 1 were synthesized.

TABLE 1 Ex. Mol In No Structure weight Mass vitro A549 NMR 1-13

522.41 295 A A (300 MHz, DMSO-d6): 1.99(4H, m), 3.11(3H, m), 3.43(2H, d,J=13.2 Hz), 6.91(2H, m), 7.20(1H, s), 7.35 (3H, m), 7.94(1H, d,J=8.5Hz), 8.46(1H, br), 8.76(1H, br), 13.20 (1H, br). 1-14

330.82 295 B B (300 MHz, DMSO-d6): 1.67-2.05 (6H, m), 3.19-3.26(1H, m),3.42- 3.51(1H, m), 4.30-4.33(1H, m), 6.91-6.95(2H, m), 7.35-7.40(1H, m),7.65(1H, br s), 8.07(1H, s), 8.18 (1H, d, J=7.3Hz), 9.41-9.45(1H, m),10.27-10.30(1H, m). 1-15

523.40 296 B B (300 MHz, DMSO-d6): 3.00(3H, m), 3.69(3H, m), 4.06(1H,m), 4.55(1H, d, J=3.7Hz), 5.14(1H, dd, J=4.0, 11.7Hz), 6.95(2H, m),7.38(1H, t, J= 8.3Hz), 7.81(3H, m), 7.98(1H, d, J=8.3Hz), 9.40(3H, br),13.5Hz, (1H, s). 1-16

378.86 343 D (500 MHz, DMSO-d6): 2.89(1H, d, J=14.8Hz), 2.92-3.19(2H,m), 3.38- 3.41(2H, m), 6.54(1H, m), 6.59 (1H, d, J=7.9Hz), 6.84-6.90(2H,m), 6.94-6.97(2H, m), 7.30-7.37 (4H, m), 7.96(1H, d, J=8.2Hz), 13.45(1H,br). 1-17

344.85 309 B B (300 MHz, DMSO-d6): 1.78-1.96 (4H, m), 2.20(3H, s),2.87-2.93(1H, m), 3.23-3.43(4H, m), 6.32(0.7H, br s), 6.82(1H, t,J=7.5Hz), 7.25 (1H, d, J=7.2Hz), 7.42(3H, br s), 7.91(1H, d, J=8.3Hz),13.88(1H, br s). 1-18

346.82 311 B B (300 MHz, DMSO-d6): 1.76-1.95(4 H, m), 2.89(1H, m),3.22(1H, m), 3.31-3.39(3H, m), 6.26(1H, d, J= 2.3Hz), 6.36(1H, dd,J=2.3, 8.7 Hz), 7.24(1H, s), 7.31(1H, br s), 7.87(1H, d, J=8.7Hz),8.77(1H, br s), 9.36(1H, br s). 1-19

426.37 313 A A (500 MHz, DMSO-d6): 1.76-1.97 (4H, m), 2.88-2.90(1H, m),3.22- 3.27(4H, m), 6.88-6.94(1H, m), 7.30-7.35(1H, m), 7.44(1H, s),7.55(2H, br s), 7.88(1H, d, J=8.5 Hz), 8.60(1H, br s), 8.99(1H, br s),13.83(1H, br s). 1-20

442.83 329 A A (500 MHz, DMSO-d6): 1.74-1.97 (4H, m), 2.86-2.92(1H, m),3.23- 3.45(4H, m), 6.94(1H, t, J=7.8Hz), 7.47(1H, br s), 7.55(1H, dd,J=7.6, 1.4Hz), 7.59(2H, br s), 8.07(1H, dd, J=8.5, 1.4Hz), 8.60(1H, brs), 8.99 (1H, br s), 14.58(1H, br s). 1-21

344.85 309 C B (500 MHz, DMSO-d6): 1.79-1.95 (4H, m), 2.20(3H, s),2.90(1H, dd, J= 10.4, 11.3Hz), 3.23-3.42(4H, m), 6.82(1H, t, J=7.6Hz),7.26(1H, d, J=7.6Hz), 7.43(2H, br s), 7.43(1H, s), 7.91(1H, d, J=7.6Hz),8.86(1H, br), 9.51(1H, br), 13.91(1H, br). 1-22

344.85 309 A A (300 MHz, DMSO-d6): 1.40-2.20 (8H, m), 2.60-2.88(1H, m),3.10 and 3.48(1H, br s), 6.75-6.95(2H, m), 7.00-7.85(3H, m), 7.85-8.25(1H, m), 8.14(1H, br s), 8.37(1H, br s). 1-23

344.85 309 A A (300 MHz, DMSO-d6): 1.27-2.17 (8H, m), 2.75-3.00(1H, m),3.19 (1H, br s), 6.70-7.00(2H, m), 7.00- 7.80(3H, m), 7.80-8.40(3H, m).1-24

344.85 309 A B (500 MHz, DMSO-d6): 1.25-1.38 (1H, m), 1.58-1.70(1H, m),1.72- 1.84(2H, m), 2.15-2.25(1H, m), 2.60-2.85(4H, m), 3.10-3.22(2H, m),6.88-6.94(2H, m), 7.34(1H, t, J=7.7Hz), 7.38(1H, s), 7.45(2H, br),7.95(1H, d, J=8.5Hz), 8.70(1H. br), 9.05(1H, br). 1-25

366.85 331 A B (300 MHz, DMSO-d6): 3.20(1H, dd, J=9.5, 13.2Hz), 3.22(1H,dd, J=5.7, 13.2Hz), 4.54(1H, br), 6.90-6.95 (2H, m), 7.10-7.13(2H, m),7.22- 7.51(7H, m), 8.09(1H, s), 8.12(1H, d, J=7.9Hz), 9.00(3H, br). 1-26

396.88 361 A B (500 MHz, DMSO-d6): 3.11-3.24 (1H, m), 3.27-3.45(1H, m),3.68 (3H, s), 5.24(1H, br s), 6.79(2H, d, J=8.8Hz), 6.88-6.99(2H, m),7.08 (2H, d, J=8.8Hz), 7.33-7.41(3H, m), 7.63(1H, s), 8.00(1H, dd, J=1.3, 8.2Hz), 10.14(1H, s). 1-27

378.86 343 C (300 MHz, DMSO-d6): 2.77-2.86 (1H, m), 4.82(1H, d, J=17.4Hz), 5.08(1H, dd, J=4.2, 11.9Hz), 5.20 (1H, d, J=17.4Hz), 6.93-7.00(2H,m), 7.28-7.51(4H, m) 7.83(1H, s), 7.94(1H, br s), 8.07(1H, dd, J=7.2,14.9Hz), 9.53(1H, s). 1-28

332.84 297 A C (300 MHz, DMSO-d6): 0.85(3H, t, J=6.7Hz), 1.17-1.30(4H,m), 1.88- 1.94(1H, m), 1.97-2.12(1H, m), 4.29(1H, br s), 6.93(2H, t,J=6.5 Hz), 7.37(1H, t, J=7.6Hz), 7.60 (1H, br s), 7.92(1H, s), 8.07(1H,dd, J=7.8, 14.5Hz), 8.99(2H, s). 1-29

458.44 345 B C (300 MHz, DMSO-d6): 2.08(1H, m), 5.03(1H, dd, J=4.1,6.8Hz), 6.92 (2H, m), 7.15-7.39(6H, m), 7.70 (3H, d, J=5.6Hz), 8.06(1H,dd, J= 1.3, 8.3Hz), 8.87(1H, br), 9.25(1H, m), 10.34(1H, m), 13.69(1H,s). 1-30

384.84 349 A B (300 MHz, DMSO-d6): 3.15-3.23 (1H, m), 3.47-3.53(1H, m),4.47- 4.51(1H, m), 6.91-6.98(2H, m), 7.06-7.20(4H, m), 7.35-7.48(1H, m),8.11(1H, s), 8.13-8.17(1H, m), 9.10(2H, br s). 1-31

316.79 281 B B (300 MHz, DMSO-d6): 1.34-1.53 (1H, m), 2.30-2.47(3H, m),3.40- 3.67(2H, m), 5.03-5.14(1H, m), 6.82-7.00(2H, m), 7.28-7.44(1H, m),7.65(1H, s), 7.97(1H, d, J=8.2 Hz), 8.10(3H, br), 9.85(1H, br s),10.02(1H, br s). 1-32

290.75 255 A B (300 MHz, DMSO-d6): 1.58(3H, d, J=6.9Hz), 4.45(1H, m),6.91-6.96 (2H, m), 7.34-7.40(1H, m), 7.90 (1H, s), 8.06(1H, d, J=8.0Hz),8.94(1H, br s). 1-33

396.37 283 A B (300 MHz, DMSO-d6): 0.64(3H, d, J=6.8Hz), 1.08(3H, d,J=6.9Hz), 4.96(1H, s), 6.90-6.95(2H, m), 7.36(1H, t, J=8.4Hz), 7.61(1H,s), 7.72(2H, br s), 8.05(1H, d, J=7.0 Hz), 10.21(1H, s), 13.65(1H, s).1-34

366.85 331 A B (300 MHz, DMSO-d6): 3.18(1H, dd, J=9.4, 13.2Hz), 3.21(1H,dd, J= 5.7, 13.2Hz), 4.54(1H, br), 6.91- 6.99(2H, m), 7.10-7.13(2H, m),7.22-7.29(4H, m), 7.35-7.55(3H, m), 8.06(1H, s), 8.12(1H, d, J=7.9 Hz),9.02(3H, br). 1-35

366.85 no peak A B (300 MHz, DMSO-d6): 3.26-3.34 (1H, m), 3.48-3.57(1H,m), 4.02- 4.80(1H, m), 6.87-6.93(2H, m), 7.30-7.43(1H., m), 7.53(2H, d,J= 6.4Hz), 7.89(1H, d, J=8.8Hz), 8.79(2H, br s). 1-36

382.35 269 A B (300 MHz, DMSO-d6): 1.21(3H, d, J=6.4Hz), 2.88-3.08(2H,m), 3.73 (1H, br s), 6.89(1H, s), 6.92(1H, br s), 7.35(2H, t, J=6.8Hz),7.43(1H, br s), 7,95(3H, br s), 13.36(1H, s). 1-37

368.32 255 A A (500 MHz, DMSO-d6): 3.03(2H, t, J=7.5Hz), 3.23(2H, t,J=7.7Hz), 6.89-6.99(2H, m), 7.35(2H, t, J= 7.1Hz), 7.41(2H, br s),7.88-7.98 (4H, m), 13.38 1H, s). 1-38

337.38 338 A B (500 MHz, DMSO-d6): 1.47(2H, dq, J=3.0, 12.8Hz), 1.72(2H,dq, J= 3.0, 12.8Hz), 1.84(2H, dd, J=3.0, 12.8Hz), 2.05(2H, dd, J=3.0,12.8 Hz), 2.33(2H, tt, J=3.3, 12.3Hz), 2.73(2H, tt, J=3.2, 12.0Hz),6.86- 6.93(2H, m), 7.23-7.38(4H, m), 8.04(1H, d, J=7.9Hz), 12.14(1H, brs), 13.51(1H, br s). 1-39

353.30 C C 1-40

506.05 470 B B (300 MHz, DMSO-d6): 1.25(1H, br), 1.60(5H, br m),2.69-2.80(4H, m), 3.04(2H, d, J=7.5Hz), 3.12-3.20 (2H, m), 5.19(1H, dd,J=7.4, 15.4 Hz), 6.91-6.97(2H, m), 7.21-7.45 (8H, m), 7.56(1H, s),7.96-7.99 (1H, m), 9.14(1H, d, J=8.1Hz), 9.58(1H, br), 13.33(1H, br s).1-41

661.61 B B 1-42

486.47 373 C B (300 MHz, DMSO-d6): 1.85(4H, m), 3.00(5H, m), 3.62(3H, d,J=11.2 Hz), 6.90(2H, m), 7.37(3H, m), 8.02(1H, dd, J=1.4, 8.4Hz), 13.4(1H, s). 1-43

506.57 393 A A (500 MHz, DMSO-d6): 0.93(6H, m), 1.68(8H, m), 2.01(4H,m), 3.00(5H, m), 6.92(2H, m), 7.35(4H, m), 8.02 (1H, d, J=13.2Hz),8.84(1H, br), 13.4(1H, s). 1-44

369.30 256 B B (300 MHz, DMSO-d6): 3.18(2H, t, J=6.2Hz), 4.76(2H, t,J=6.2Hz), 6.90(2H, m), 7.23-7.82(4H, m) 7.95 (1H, dd, J=1.6, 8.4Hz),13.39(1H, s).

Example 2-1

(1) With the use of starting compound 1E instead of 1A, tert-butyl3-{2-amino-6-[2,6-bis(benzyloxy)phenyl]-3-cyano-4-pyridinyl}-1-piperidinecarboxylate(3.54 g, yield; 30%) was prepared in a similar manner as described instep (1) in Example 1-1.

(2) A suspension of tert-butyl3-{2-amino-6-[2,6-bis(benzyloxy)phenyl]-3-cyano-4-pyridinyl}-1-piperidinecarboxylate(3.46 g, 5.9 mmol) and Pd/C (10%, 600 mg) in ethyl acetate (50 mL)including acetic acid (5 mL) was stirred for 2 days at room temperatureunder a hydrogen atmosphere. The reaction mixture was diluted with THF(200 mL) and filtered through Celite®. The filtrate was concentratedunder reduced pressure. The residue was washed with ethyl acetate andhexane, and dried under reduced pressure to give tert-butyl3-[2-amino-3-cyano-6-(2,6-dihydroxy-phenyl)-4-pyridinyl]-1-piperidinecarboxylate(1.82 g, yield; 76%).

(3) tert-butyl3-[2-amino-3-cyano-6-(2,6-dihydroxyphenyl)-4pyridinyl]-1-piperidinecarboxylate(200 mg, 0.49 mmol) was treated under acidic conditions in a similarmanner as described in the step (3) of Example 1-1. The resultingprecipitate was collected by filtration, washed with ethyl acetate andhexane, and dried under reduced pressure to give2-amino-6-(2,6-dihydroxyphenyl)-4-(3-piperidinyl)nicotinonitrilehydrochloride (123 mg, yield; 73%).

Molecular weight: 346.82; Mass spectrometry: 311 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 1.66-1.99 (5H, m), 2.91-3.20 (2H, m), 3.23-3.42 (3H, m), 6.41(2H, d, J=7.9 Hz), 7.08 (1H, t, J=7.9 Hz), 7.28 (2H, br d), 7.65 (1H,s), 8.82 (1H, br d), 9.17 (1H, br d).

Example 2-2

A suspension of tert-butyl3-[2-amino-3-cyano-6-(2,6-dihydroxyphenyl)-4-pyridinyl]-1-piperidinecarboxylate(300 mg, 0.73 mmol) obtained in the step (2) of Example 2-1, benzylbromide (0.1 mL, 0.80 mmol) and potassium carbonate (303 mg, 2.19 mmol)in a solution of acetone (15 mL) and THF (15 mL) was stirred for 2 daysat room temperature. The reaction mixture was concentrated under reducedpressure, then dissolved in ethyl acetate (100 mL) and water (100 mL).The separated aqueous phase was extracted with ethyl acetate (100 mL×2).The combined organic phase was washed with brine (100 mL), dried overNa₂SO₄, filtered, and concentrated under reduced pressure. The crudeproduct was purified by column chromatography on silica gel(hexane/ethyl acetate, 4:1-2:1-1:1) to give tert-butyl3-{2-amino-6-[2-(benzyloxy)-6-hydroxyphenyl]-3-cyano-4-pyridinyl}-1-piperidinecarboxylate(239 mg, yield; 65%).

To a solution of tert-butyl3-{2-amino-6-[2-(benzyloxy)-6-hydroxyphenyl]-3-cyano-4-pyridinyl}-1-piperidinecarboxylate(239 mg, 0.48 mmol) in dioxane (25 mL) was added 4N HCl/dioxane (25 mL).The reaction mixture was stirred at room temperature overnight. Theresulting precipitate was collected by filtration, washed with ethylacetate and hexane, dried under reduced pressure to give2-amino-6-[2,-(benzyloxy)-6-hydroxyphenyl]-4-(3-piperidinyl)nicotinonitrilehydrochloride (184 mg, yield; 88%).

Molecular weight: 436.95; Mass spectrometry: 401 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 1.07-1.14 (1H, m), 1.65-1.74 (3H, m), 2.57-2.73 (2H, m),3.07-3.12 (1H, m), 3.17-3.23 (2H, m), 5.07 (1H, d, J=11.3 Hz), 5.12 (1H,d, J=11.3 Hz), 6.57 (1H, d, J=8.3 Hz), 6.73 (1H, d, J=8.3 Hz), 7.24-7.49(8H, m), 8.72 (1H, br d), 9.01 (1H, br d), 12.54 (1H, s).

Example 2-3

With the use of the starting compound 1G instead of 1A and4-formyl-piperidine-1-carboxylic acid tert-butyl ester prepared in asimilar manner as that of the starting compound 2B, tert-butyl4-(2-amino-3-cyano-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-4-pyridinyl)-1-piperidinecarboxylatewas prepared in a similar manner as that of step (1) of Example 1-1.

To a stirred solution of tert-butyl4-(2-amino-3-cyano-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-4-pyridinyl)-1-piperidinecarboxylate(0.63 g, 1.077 mmol) in 1,4-dioxane (25 mL) was added 4N HCl in1,4-dioxane (25 mL). The mixture was stirred at room temperature for 12hrs. The resulting solid was collected with filtration and washed withdiisopropyl ether to give2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-(3-piperidinyl)nicotinonitrilehydrochloride as a yellow solid (0.340 g, yield; 79%).

Molecular weight: 400.91; Mass spectrometry: 365 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)A; ¹H-NMR (500 MHz,DMSO-d6): 0.31-0.34 (2H, m), 0.55-0.59 (2H, m), 1.03-1.04 (1H, m),1.91-1.99 (4H, m), 3.06-3.14 (3H, m), 3.37-3.39 (2H, m), 3.87 (2H, d,J=7.3 Hz), 6.55 (2H, dd, J=9.9, 8.2 Hz), 7.23 (1H, t, J=8.2 Hz),9.19-9.11 (1H, m), 9.20-9.23 (1H, m).

Examples 2-4 to 2-87

According to the similar synthetic procedure of Examples 2-1 to 2-3,compounds shown in Tables 2 were prepared.

TABLE 2 Ex. Mol In No Structure weight Mass vitro A549 NMR 2-04

346.82 311 (500 MHz, DMSO-d6): 1.88-1.98 (4H, m), 3.05-3.10(3H, m),3.38- 3.57(2H, m), 6.43(2H, d, J=8.2Hz), 7.09(1H, t, J=8.2Hz),7.56-7.60(1H, brs), 7.60(1H, s), 8.94-9.01(2H, m). 2-05

505.84 470 A B (500 MHz, DMSO-d6): 1.26(1H, br), 1.75(3H, m), 2.71(1H,br), 2.88(1H, br), 3.18(1H, br), 3.28(1H, br), 5.07 (1H, d, J=11.7Hz),5.12(1H, d, J= 11.7Hz), 6.59(1H, m), 6.69(1H, d, J=8.5Hz), 7.15(1H, d,J=8.2Hz), 7.26 (2H, m), 7.45(1H, dd, J=1.9, 8.2Hz), 7.69(1H, d,J=8.2Hz), 8.76(1H, br), 8.92(1H, br), 9.09(1H, br), 9.40(1H, br # s).2-06

528.54 415 A B (500 MHz, DMSO-d6): 0.92(1H, ddd, J=3.5, 3.5, 12.3Hz),1.58-1.72 (3H, m), 2.27(3H, s), 2.30-2.70(3H, m), 3.06(1H, t, J=12.0Hz),3.25(1H, m), 5.08(1H, d, J=11.0Hz), 5.13(1H, d, J=10.8Hz), 6.56(1H, d,J=8.2Hz), 6.77(1H, d, J=8.2Hz), 7.23-7.31(6H, m), 7.43(1H, d, J=7.6Hz),8.59(1H, br), 8.74(1H, br), 12.82(1H, br). 2-07

528.54 415 A B (500 MHz, DMSO-d6): 0.92(1H, ddd, J=3.8, 4.1, 12.6Hz),1.58-1.72 (3H, m), 2.27(3H, s), 2.30-2.70(3H, m), 3.07(1H, m), 3.25(1H,t, J=9.5 Hz), 5.08(1H, d, J=10.7Hz), 5.13(1H, d, J=10.7Hz), 6.56(1H, dd,J=1.0, 8.2 Hz), 6.77(1H, d, J=8.2Hz), 7.23- 7.31(6H, m), 7.43(1H, d,J=7.3Hz), 8.59(1H, br), 8.75(1H, br), # 12.83 (1H, br). 2-08

528.54 415 A A (500 MHz, DMSO-d6): 0.85(1H, m), 1.62-1.95(3H, m),2.31(3H, s), 2.30- 2.70(3H, m), 3.08(1H, m), 3.20-3.50 (1H, m), 5.02(1H,d, J=10.7Hz), 5.07 (1H, d, J=10.7Hz), 6.55(1H, dd, J= 1.0, 8.2Hz),6.72(1H, d, J=8.2Hz), 7.22-7.27(6H, m), 7.37(1H, d, J= 7.9Hz), 8.58(1H,br), 8.76(1H, br), 12.68(1H, br). 2-09

582.51 469 A B (500 MHz, DMSO-d6): 1.17-1.24 (1H, m), 1.64-1.72(3H, m),2.30- 2.50(2H, m), 2.63-2.68(1H, m), 2.81 (1H, br t, J=11.7Hz), 3.12(1H,m), 5.17(1H, d, J=11.7Hz), 5.22(1H, d, J=11.7Hz), 6.58(1H, d, J=8.2Hz),6.71(1H, d, J=8.2Hz), 7.11(1H, s), 7.17(2H, br s), 7.25(1H, t, J=8.2Hz),7.64(1H, dd, J=7.9, 8.2Hz), 7.71 # (2H, m), 7.75(1H, d, J=7.9Hz), 8.60(1H, br), 8.80(1H, br), 11.9(1H, br). 2-10

582.51 469 A B (500 MHz, DMSO-d6): 1.24-1.31 (1H, m), 1.71(3H, m),2.60(2H, m), 2.69(1H, m), 2.88(1H, br t, J=12.3 Hz), 3.13(1H, m),5.19(1H, d, J=12.3 Hz), 5.24(1H, d, J=12.3Hz), 6.57(1H, d, J=8.2Hz),6.70(1H, d, J=8.2Hz), 7.17(1H, s), 7.19(2H, br s), 7.24(1H, t, J=8.2Hz),7.66(2H, d, J=7.9Hz), 7.77(2H, d, J=7.9Hz), 8.66(2H, # br), 11.95(1H,s). 2-11

593.40 no peak A B (500 MHz, DMSO-d6): 1.22(1H, m), 1.74(3H, m),2.69-2.81(2H, m), 3.13 (1H, t, J=12.3Hz), 5.07(1H, d, J= 11.4Hz),5.12(1H, d, J=11.4Hz), 6.57 (1H, d, J=8.5Hz), 6.69(1H, d, J=8.5 Hz),7.16(1H, s), 7.19(2H, br s), 7.25 (1H, t, J=8.5Hz), 7.37(1H, t, J=7.9Hz), 7.46(1H, d, J=7.9Hz), 7.55(1H, d, J=7.9Hz), 7.61 # (1H, br s), 8.59(1H, br s), 8.79(1H, br s), 12.10(1H, br). 2-12

559.51 446 A B (500 MHz, DMSO-d6): 1.24(1H, m), 1.73(3H, m),2.30-2.70(3H, m), 2.80 (1H, m), 3.14(1H, m), 5.42(1H, d, J= 13.2Hz),5.46(1H, d, J=13.2Hz), 6.58 (1H, d, J=8.2Hz), 6.67(1H, d, J=8.2 Hz),7.10(1H, s), 7.19(2H, br s), 7.24 (1H, t, J=8.2Hz), 7.64(1H, dt, J=1.3,8.2Hz), 7.71(1H, d, J=7.6Hz), 7.78 (1H, d, J=7.6Hz), 8.11(2H, d, # J=8.2Hz), 8.56(1H, br), 8.74(1H, br), 11.94(1H, br). 2-13

559.51 446 A B (500 MHz, DMSO-d6): 1.29(1H, m), 1.71(3H, m),2.30-2.50(2H, m), 2.69 (1H, m), 2.84(1H, dd, J=11.4, 11.6 Hz), 3.14(1H,m), 5.23(1H, d, J= 12.3Hz), 5.28(1H, d, J=12.3Hz), 6.59 (1H, dd, J=1.0,8.5Hz), 6.71(1H, d, J=8.5Hz), 7.12(1H, s), 7.16(2H, br s), 7.25(1H, t,J=8.5Hz), 7.71(1H, t, J=7.9Hz), 7.89(1H, d, J=7.6Hz), # 8.20(1H, dd,J=1.3, 8.2Hz), 8.24(1H, s), 8.58(1H, br), 8.82(1H, br). 2-14

559.51 446 A A (500 MHz, DMSO-d6): 1.38(1H, m), 1.75(3H, m),2.30-2.50(1H, m), 2.73 (1H, dd, J=10.7, 12.0Hz), 2.88(1H, dd, J=10.7,11.6Hz), 3.14(1H, m), 3.29(1H, m), 5.27(2H, dd, J=12.9, 16.1Hz),6.58(1H, dd, J=1.0, 8.2Hz), 6.69(1H, d, J=8.5Hz), 7.13(1H, s), 7.19(2H,br s), 7.24(1H, t, J=8.2 Hz), 7.69(1H, t, J=8.8Hz), 8.26(1H, d, J= #8.8Hz), 8.54(1H, s), 8.79(1H, br), 11.8(1H, br). 2-15

539.52 426 A A (500 MHz, DMSO-d6): 1.24(1H, m), 1.68-1.81(3H, m),2.30-2.50(2H, m), 2.72(1H, dd, J=10.1, 12.3Hz), 2.82(1H, dd, J=10.4,12.0Hz), 3.13 (1H, m), 5.12(1H, d, J=11.7Hz), 5.18 (1H, d, J=11.7Hz),6.58(1H, d, J= 8.5Hz), 6.71(1H, d, J=8.5Hz), 7.13 (1H, s), 7.18(2H, brs), 7.26(1H, t, J=8.5Hz), 7.62(1H, t, J=7.9Hz), # 7.79 (1H, d, J=7.9Hz),7.84(1H, d, J= 7.9Hz), 7.88(1H, s), 8.59(1H, br), 8.76(1H, br),11.95(1H, br). 2-16

539.52 426 A A (500 MHz, DMSO-d6): 1.31(1H, m), 1.68-1.91(3H, m),2.30-2.50(2H, m), 2.73(1H, m), 2.87(1H, dd, J=11.4, 12.3Hz), 3.14(1H, t,J=12.3Hz), 5.19 (1H, d, J=12.6Hz), 5.22(1H, d, J= 12.6Hz), 6.57(1H, d,J=8.2Hz), 6.68 (1H, d, J=8.2Hz), 7.13(1H, s), 7.18 (2H, br s), 7.24(1H,t, J=8.2Hz), 7.62 (1H, t, J=8.2Hz), 7.88(1H, d, # J= 8.2Hz), 8.58(1H,br), 8.80(1H, br), 11.84(1H, br s). 2-17

548.95 435 A B (500 MHz, DMSO-d6): 1.06(1H, m), 1.60-1.80(3H, m),2.50-2.70(m, 4H), 3.10(1H, m), 5.13(1H, d, J=11.4 Hz), 5.21(1H, d,J=11.4Hz), 6.58(1H, dd, J=0.9, 8.2Hz), 6.74(1H, d, J=8.2 Hz), 7.21(1H,s), 7.24(2H, br), 7.28 (1H, t, J=8.2Hz), 7.41(1H, dt, J=1.6, 7.3Hz),7.44(1H, dt, J=1.9, 7.6Hz), 7.57(1H, dd, J=1.6, 7.9Hz), # 7.61 (1H, dd,J=1.9, 7.3Hz), 8.61(1H, br), 8.77(1H, br). 2-18

548.95 435 A B (500 MHz, DMSO-d6): 1.22(1H, m), 1.74(3H, m), 2.71(2H,m), 3.13(1H, t, J=12.3Hz), 5.08(1H, d, J=11.3Hz), 5.13(1H, d, J=11.3Hz),6.57(1H, d, J=8.2Hz), 6.70(1H, d, J=8.2Hz), 7.17 (1H, s), 7.20(2H, br),7.25(1H, t, J= 8.2Hz), 7.42(3H, m), 7.48(1H, s), 8.62(1H, br), 8.82(1H,br). 2-19

548.95 435 A A (500 MHz, DMSO-d6): 1.13(1H, m), 1.73(3H, m),2.50-2.80(3H, m), 3.11 (1H, t, J=12.3Hz), 5.07(1H, d, J= 11.0Hz),5.12(1H, d, J=11.0Hz), 6.56 (1H, d, J=8.2Hz), 6.70(1H, d, J=8.2 Hz),7.19(1H, s), 7.21(2H, br), 7.25 (1H, t, J=8.2Hz), 7.49(4H, dt, J=2.5,8.8Hz), 8.61(1H, br), 8.80(1H, br), 12.33(1H, br). 2-20

583.40 470 A B (500 MHz, DMSO-d6): 0.85(1H, m), 1.53(1H, d, J=12.6Hz),1.69(2H, m), 3.07(1H, m), 5.24(1H, d, J=10.4Hz), 5.33(1H, d, J=10.4Hz),6.60(1H, dd, J=0.9, 8.2Hz), 6.84(1H, d, J=8.5Hz), 7.14(1H, s), 7.29(2H,br), 7.32(1H, t, J=8.2Hz), 7.51(4H, dd, J=7.6, 8.8 Hz), 7.63(2H, d,J=7.9Hz), 8.64(1H, br), 8.77(1H, br), 12.95(1H, br). 2-21

564.57 451 A A (500 MHz, DMSO-d6): 0.82(1H, m), 1.09(1H, m), 1.41(2H,m), 2.08(1H, m), 2.99(2H, m), 5.24(1H, d, J=10.7 Hz), 5.30(1H, d,J=10.7Hz), 6.57(1H, dd, J=1.0, 8.2Hz), 6.80(1H, d, J=8.2 Hz), 7.23(2H,br), 7.26(1H, s), 7.29 (1H, t, J=8.2Hz), 7.55(2H, m), 7.62 (1H, dd,J=1.6, 8.2Hz), 7.95(3H, m), 8.03(1H, s), 8.55(1H, br), 12.65(1H, s).2-22

590.61 477 A B (500 MHz, DMSO-d6): 0.94(1H, m), 1.63(3H, m), 3.07(1H, t,J=11.4Hz), 4.90(1H, d, J=10.4Hz), 4.99(1H, d, J=10.4Hz), 6.51(1H, d,J=8.2Hz), 6.53(1H, d, J=8.2Hz), 7.19(1H, t, J= 8.2Hz), 7.26(2H, br),7.35(6H, m), 7.48(2H, m), 7.66(1H, d, J=7.3Hz), 8.60(1H, br), 8.76(1H,br). 2-23

436.95 401 A B (500 MHz, DMSO-d6): 1.65-1.89 (4H, m), 2.93-3.10(3H, m),3.29(2H, d, J=12.3Hz), 5.13(2H, s), 6.62(1H, d, J=8.2Hz), 6.72(1H, d,J=8.2Hz), 7.13(1H, s), 7.28(1H, t, J=8.2Hz), 7.33(1H, t, J=7.4Hz),7.40(2H, t, J= 7.4Hz), 7.46(2H, t, J=7.4Hz), 7.72 (2H, br), 9.10(1H, brs). 2-24

593.40 480 A A (500 MHz, DMSO-d6): 1.14(1H, m), 1.74(3H, m), 2.69(2H,m), 3.11(1H, t, J=12.0Hz), 5.05(1H, d, J=11.4Hz), 5.10(1H, d, J=11.4Hz),6.56(1H, d, J=8.2Hz), 6.70(1H, d, J=8.2Hz), 7.19 (1H, s), 7.22(2H, br),7.25(1H, t, J= 8.2Hz), 7.44(2H, d, J=8.2Hz), 7.61 (2H, d, J=8.2Hz),8.60(1H, br), 8.81 (1H, br), 12.36(1H, br). 2-25

532.50 419 A A (500 MHz, DMSO-d6): 1.09(1H, m), 1.71(3H, m), 2.64(2H,m), 3.10(1H, t, J=12.3Hz), 3.28(2H, t, J=10.4Hz), 5.13(1H, d, J=11.0Hz),5.18(1H, d, J=11.0Hz), 6.58(1H, d, J=8.5Hz), 6.76(1H, d, J=8.5Hz),7.20(1H, s), 7.26(4H, m), 7.47(1H, m), 7.58(1H, t, J=7.6Hz), 8.63(1H,br), 8.82(1H, br). 2-26

550.49 A A (500 MHz, DMSO-d6): 1.09(1H, m), 1.71(3H, m), 2.64(2H, m),3.10(1H, t, J=12.3Hz), 3.28(2H, t, J=10.4Hz), 5.13(1H, d, J=11.0Hz),5.18(1H, d, J=11.0Hz), 6.58(1H, d, J=8.5Hz), 6.76(1H, d, J=8.5Hz),7.20(1H, s), 7.26(4H, m), 7.47(1H, m), 7.58(1H, t, J=7.6Hz), 8.63(1H,br), 8.82(1H, br). 2-27

532.50 419 A A (500 MHz, DMSO-d6): 1.25(1H, m), 1.73(3H, m), 2.69(1H,dd, J=10.8, 11.7Hz), 2.78(1H, dd, J=11.7, 11.7 Hz), 3.13(1H, t,J=12.3Hz), 5.08(1H, d, J=11.7Hz), 5.14(1H, d, J=11.7 Hz), 6.57(1H, d,J=8.2Hz), 6.70(1H, d, J=8.2Hz), 7.26(6H, m), 7.45(1H, m), 8.60(1H, br),8.81(1H, br), 12.19 (1H, br). 2-28

532.50 419 A A (500 MHz, DMSO-d6): 1.14(1H, m), 1.71(3H, m), 2.69(3H,m), 3.10(1H, t, J=12.3Hz), 5.06(1H, d, J=11.0Hz), 5.10(1H, d, J=11.0Hz),6.56(1H, d, J=8.5Hz), 6.72(1H, d, J=8.5Hz), 7.21 (1H, s), 7.25(5H, m),7.53(2H, dd, J= 5.7, 8.5Hz), 8.59(1H, br), 8.81(1H, br). 2-29

550.49 437 A B (500 MHz, DMSO-d6): 1.28(1H, m), 1.73(3H, m), 2.69(1H,m), 2.84(1H, dd, J=11.4, 11.4Hz), 3.13(1H, t, J= 12.3Hz), 5.05(1H, d,J=11.4Hz), 5.10 (1H, d, J=11.4Hz), 6.57(1H, d, J= 8.2Hz), 6.69(1H, d,J=8.2Hz), 7.17 (1H, s), 7.20(2H, br), 7.25(1H, t, J= 8.2Hz), 7.33(1H,br), 7.50(2H, m), 8.60(1H, br), 8.82(1H, br), # 12.07 (1H, br). 2-30

572.55 459 A A (500 MHz, DMSO-d6): 1.22(1H, m), 1.70(3H, m), 2.65(1H,m), 2.79(1H, dd, J=11.4, 11.4Hz), 3.12(1H, t, J= 12.3Hz), 3.29(2H, br),5.17(1H, d, J= 12.0Hz), 5.22(1H, d, J=12.0Hz), 6.57 (1H, d, J=8.2Hz),6.69(1H, d, J=8.2 Hz), 7.17(1H, s), 7.22(2H, br), 7.25 (1H, t, J=8.2Hz),7.59(2H, d, J=8.2 Hz), 7.99(2H, d, J=8.2Hz), 8.58 # (1H, br), 8.80(1H,br). 2-31

570.62 457 A A (500 MHz, DMSO-d6): 1.67-1.78 (3H, m), 2.71(1H, dd,J=10.4, 11.4 Hz), 2.89(1H, dd, J=11.4, 11.7Hz), 3.13(1H, t, J=12.3Hz),5.04(1H, d, J= 11.4Hz),5.10(1H, d, J=11.4Hz), 6.55 (1H, d, J=8.2Hz),6.72(1H, d, J=8.2 Hz), 7.22(2H, br s), 7.24(1H, t, J=8.2 Hz), 7.27(1H,s), 7.37(2H, d, J=8.2 Hz), 7.41(2H, d, J=8.2Hz), 8.61 # (1H, br),8.84(1H, br), 12.20(1H, br). 2-32

564.57 451 A B (500 MHz, DMSO-d6): 0.24(1H, m), 1.13(1H, d, J=13.9Hz),1.42-1.52 (2H, m), 2.04(1H, dd, J=11.0, 12.0 Hz), 2.19(1H, dd, J=11.0,12.0Hz), 2.87(1H, t, J=12.0Hz), 3.08(1H, d, J=11.4Hz), 3.15(1H, d,J=12.0Hz), 5.54(1H, d, J=11.0Hz), 5.59(1H, d, J=11.0Hz), 6.60(1H, d,J=8.2Hz), 6.95(1H, d, J=8.2Hz), 7.10(1H, s), 7.23 # (2H, br s), 7.34(1H,t, J=8.2 Hz), 7.56(3H, m), 7.71(1H, d, J=6.9 Hz), 8.00(3H, m), 8.46(1H,br), 8.60 (1H, br). 2-33

590.61 477 A B (500 MHz, DMSO-d6): 1.17(1H, m), 1.59-1.68(3H, m),2.76(1H, dd, J= 11.4, 11.7Hz), 3.07-3.17(3H, m), 5.13(1H, d, J=11.4Hz),5.18(1H, d, J=11.4Hz), 6.57(1H, d, J=8.2Hz), 6.76(1H, d, J=8.2Hz),7.24-7.94(4H, m), 7.39(1H, t, J=7.6Hz), 7.48(1H, t, J=7.6Hz), 7.56(2H,d, J=8.2Hz), 7.69 (2H, d, J=7.9Hz), 7.73(2H, d, J= # 7.9 Hz), 8.55(1H,br d), 8.77(1H, br d), 12.47(1H, br). 2-34

583.40 470 A B (500 MHz, DMSO-d6): 1.08(1H, m), 1.64-1.78(3H, m),3.12(1H, t, J=12.3 Hz), 3.32(2H, t, J=12.9Hz), 5.11(1H, d, J=11.4Hz),5.20(1H, d, J=11.4Hz), 6.59(1H, d, J=8.2Hz), 6.73(1H, d, J= 8.2Hz),7.16(1H, s), 7.25(2H, br), 7.27(1H, t, J=8.2Hz), 7.51(1H, dd, J=1.9,8.2Hz), 7.62(1H, d, J=8.2Hz), 7.76(1H, d, J=1.9Hz), 8.63 # (1H, br),8.81(1H, br). 2-35

450.97 415 A A (500 MHz, DMSO-d6): 1.09(1H, m), 1.68(3H, m), 2.32(3H,s), 3.08-3.31 (4H, m), 5.02(1H, d, J=10.6Hz), 5.07 (1H, d, J=10.6Hz),6.56(1H, d, J=8.3 Hz), 6.72(1H, d, J=8.3Hz), 7.22- 7.29(6H, m), 7.36(2H,d, J=7.5Hz), 8.79(1H, br), 9.16(1H, br). 2-36

504.94 469 A A (500 MHz, DMSO-d6): 1.29(1H, m), 1.72(3H, m), 2.92(1H,m), 3.17-3.37 (3H, m), 5.18(1H, d, J=12.4Hz), 5.24 (1H, d, J=12.4Hz),6.59(1H, dd, J= 2.3, 8.3Hz), 6.70(1H, d, J=8.3Hz), 7.18(1H, d, J=2.3Hz),7.25(1H, dt, J=7.9, 8.3Hz), 7.66(2H, d, J=8.3Hz), 7.78(2H, d, J=8.3Hz),8.78(1H, br), 9.21(1H, br). 2-37

454.94 419 A A (500 MHz, DMSO-d6): 1.15-1.20 (1H, m), 1.66-1.77(3H, m),2.52- 2.73(2H, m), 3.15-3.28(3H, m), 5.14 (1H, d, J=11.0Hz), 5.18(1H, d,J= 11.0Hz), 6.62(1H, d, J=8.2Hz), 6.78 (1H, d, J=8.2Hz), 7.20(1H, s),7.26- 7.33(3H, m), 7.46(2H, m), 7.58(2H, dt, J=1.6, 7.6Hz), 9.00(1H, brd), 9.50(1H, br d). 2-38

494.98 459 A A (500 MHz, DMSO-d6): 1.29(1H, m), 1.73(3H, m), 2.64(1H,m), 2.89(1H, dd, J=1.4, 12.7Hz), 3.24(3H, m), 3.86(3H, s), 5.17(1H, d,J=12.0Hz), 5.22(1H, d, J=12.0Hz), 6.62(1H, d, J=8.2Hz), 6.71(1H, d,J=8.5Hz), 7.20 (1H, s), 7.27(1H, dd, J=8.2, 8.5Hz), 7.59(2H, d,J=8.2Hz), 7.99(2H, d, J= 8.2Hz), 8.95(1H, br d), 9.41(1H, br d). 2-39

515.84 479 A A (500 MHz, DMSO-d6): 1.12(1H, m), 1.73(3H, m), 2.66(1H,m), 2.75(1H, t, J=12.3Hz), 3.13(1H, m), 5.05(1H, d, J=11.0Hz), 5.10(1H,d, J=11.0Hz), 6.56(1H, d, J=8.2Hz), 6.70(1H, d, J= 8.2Hz), 7.19(1H, s),7.21(2H, br s), 7.25(1H, t, J=8.2Hz), 7.44(2H, d, J= 8.5Hz), 7.63(2H, d,J=8.5Hz), 9.03 (2H, br), 12.37(1H, s). 2-40

474.44 361 A A (500 MHz, DMSO-d6): 2.92(4H, br s), 5.15(2H, s), 6.55(1H,d, J=7.2Hz), 6.68(1H, d, J=8.5Hz), 7.22-7.25(3H, m), 7.30(1H, s),7.33-7.35(1H, m), 7.40-7.45(4H, m), 7.95(1H, br s). 2-41

450.97 415 A A (500 MHz, DMSO-d6): 1.63(1H, m), 1.78-1.90(3H, m),2.85(1H, m), 3.03 (2H, dd, J=6.6, 6.9Hz), 3.12(1H, m), 3.26-3.36(3H, m),4.20(2H, dd, J= 6.6, 6.9Hz), 6.62(1H, d, J=8.2Hz), 6.63(1H, d, J=8.5Hz),7.18-7.24(5H, m), 7.23(1H, s), 7.26(1H, dd, J=8.2, 8.5Hz), 7.95(1H, brs), 9.11(1H, br s), 9.65(1H, br s). 2-42

437.93 402 A B (500 MHz, DMSO-d6): 1.42-1.86 (4H, m), 2.80-3.29(5H, m),5.27- 5.36(2H, m), 6.64(1H, d, J=8.2Hz), 6.72(1H, d, J=8.5Hz),7.26-7.32(2H, m), 7.51-7.73(3H, m), 8.15-8.18 (1H, t, J=7.5Hz),8.77-8.93(2H, m), 9.47(1H, br d, J=10.4 Hz). 2-43

437.93 402 A A (500 MHz, DMSO-d6): 1.33-1.36 (2H, m), 1.73-1.82(4H, m),2.80- 3.27(3H, m), 5.30(2H, s), 6.64(1H, d, J=8.2Hz), 6.74(1H, d,J=8.5Hz), 7.13 (1H, s), 7.29(1H, t, J=8.2Hz), 7.48 (1H, br s), 7.98(1H,m), 8.46(1H, d, J=7.6Hz), 8.84(1H, d, 5.4Hz), 8.89- 8.91(1H, m),8.95(1H, s), 9.45(1H, br d, J=10.1Hz). 2-44

437.93 402 A A (500 MHz, DMSO-d6): 1.52-1.86 (3H, m), 2.79-3.33(6H, m),5.42(2H, s), 6.63(1H, d, J=8.2Hz), 6.65(1H, d, J=8.5Hz), 7.10(1H, s),7.22-7.38(3H, m), 7.90(2H, d, J=5.7Hz), 8.82-8.89 (3H, m), 9.42(1H, brd, J=10.4Hz). 2-45

487.99 452 A A (500 MHz, DMSO-d6): 1.27-1.29 (1H, m), 1.49-1.67(3H, m),2.90- 3.30(4H, m), 4.61(2H, br s), 5.41- 5.42(2H, m), 6.63(1H, d,J=8.2Hz), 6.75(1H, d, J=8.5Hz), 7.25-7.29(2H, m), 7.66-7.71(2H, m),7.85(1H, t, J= 7.6Hz), 8.08(2H, t, J=9.3Hz), 8.57 (1H, d, J=8.5Hz),8.81-8.83(1H, m), 9.29(1H, br d, J=9.8Hz). 2-46

618.66 505 A B (500 MHz, DMSO-d6): 1.53-1.66 (3H, m), 1.81(1H, m),3.02(1H, m), 3.15-3.42(4H, m), 3.73-3.81(2H, m), 3.90-4.00(2H, m),4.04(1H, m), 6.40(1H, d, J=8.2Hz), 6.51(1H, d, J= 8.5Hz), 7.14(1H, dd,J=8.2, 8.5Hz), 7.19-7.28(10H, m), 7.36(1H, s), 8.60 (1H, br s), 8.83(1H,br s). 2-47

527.48 490 A B (500 MHz, DMSO-d6): 0.76-0.86 (1H, m), 1.38(1H, br d,J=12.3Hz), 3.05(1H, t, J=10.4Hz), 3.81(2H, dd, J=12.0, 29.6Hz), 5.39(2H,dd, J= 11.7, 40.0Hz), 6.60(1H, d, J=8.2Hz), 6.86(1H, d, J=8.5Hz),7.15(1H, s), 7.29-7.44(4H, m), 7.79(1H, d, J= 1.9Hz), 7.99(1H, s),8.07(1H, d, J= 8.8Hz), 8.70-8.80(1H, m), 9.07 # (1H, d, J=10.7Hz). 2-48

506.45 393 A A (500 MHz, DMSO-d6): 1.62-1.95 (4H, m), 2.87-3.21(5H, m),5.41(2H, s), 6.60(1H, d, J=8.2Hz), 6.76(1H, d, J=8.5Hz), 7.26-7.30(2H,m), 7.46 (1H, s), 8.61-8.63(1H, m), 8.85(1H, br d, J=10.4Hz), 9.74(1H,s). 2-49

519.48 406 A A (500 MHz, DMSO-d6): 1.52-1.91 (4H, m), 2.42(3H, s),2.87-3.39(5H, m), 5.12-5.26(2H, m), 6.31(1H, s), 6.58(1H, d, J=8.2Hz),6.71(1H, d, J= 8.5Hz), 7.24-7.28(3H, m), 7.36(1H, s), 8.63-8.65(1H, m),8.90-8.93(1H, br d, J=10.4Hz). 2-50

465.00 429 A A (500 MHz, DMSO-d6): 1.64-1.76 (3H, m), 1.90(1H, m),2.03-2.08(2H, m), 2.67(2H, t, J=7.6Hz), 2.78(1H, m), 3.06(1H, m),3.21-3.27(2H, m), 3.35(1H, m), 3.98(12H, ddd, J=4.1, 6.3, 9.5Hz),6.53(1H, d, J=8.2Hz), 6.54(1H, d, J=8.2Hz), 7.15-7.28(5H, m), 7.21(1H,t, J=8.2Hz), 7.31(1H, br s), 7.33(1H, s), 8.74(1H, br s), 9.10 # (1H, brs). 2-51

464.96 429 A A (500 MHz, DMSO-d6): 1.91(3H, m), 3.03(1H, m), 5.68(2H,s), 6.58(1H, d, J=8.3Hz), 6.76(1H, d, J=8.3Hz), 7.27 (1H, t, J=8.3Hz),7.37(1H, br), 7.60 (2H, t, J=7.2Hz), 7.73(1H, t, J=7.2 Hz), 8.11(2H, d,J=7.2Hz), 8.18(1H, br), 8.92(1H, br), 9.34(1H, br). 2-52

450.97 415 A A (300 MHz, DMSO-d6): 1.91-1.92 (4H, m), 3.07-3.11(2H, m),3.09(3H, t, J=7.2Hz), 3.36-3.40(2H, m), 4.22 (2H, t, J=7.2Hz), 6.55(1H,d, J=8.3 Hz), 6.61(1H, d, J=7.9Hz), 7.20(1H, dd, J=7.9, 8.3Hz),7.22-7.24(5H, m), 7.25(1H, s), 7.28(1H, br s), 8.86(2H, br s). 2-53

442.99 407 A A (300 MHz, DMSO-d6): 1.09-1.17 (2H, m), 1.21-1.25(4H, m),1.66- 1.75(5H, m), 1.81-1.96(3H, m), 2.84-2.99(2H, m), 3.03-3.16(2H, m),3.31-3.48(2H, m), 3.67-3.85 (2H, m), 6.55(1H, d, J=7.9Hz), 6.57 (1H, d,J=8.3Hz), 7.22(1H, dd, J=7.9, 8.3Hz), 7.27(1H, s), 7.45(1H, br s),8.98(1H, br s), 9.44(1H, br s). 2-54

388.90 353 A A (300 MHz, DMSO-d6): 1.29(6H, d, J=6.0Hz), 1.61-1.95(4H,m), 2.88- 3.10(2H, m), 3.28-3.49(3H, m), 4.65 (1H, sep, J=6.0Hz),6.51(1H, d, J= 7.9Hz), 6.60(1H, d, J=8.7Hz), 7.22 (1H, dd, J=7.9,8.7Hz), 7.33(1H, s), 7.42(1H, br s), 8.87(1H, br s), 9.24 (1H, br s).2-55

388.90 353 A A (500 MHz, DMSO-d6): 0.94(3H, t, J=7.3Hz), 1.67(1H, m),1.78(1H, m), 1.76-1.85(2H, m), 1.91-1.99(2H, m), 2.88(1H, m), 3.06(1H,m), 3.27(1H, m), 3.30-3.32(3H, m), 3.38(1H, m), 3.96(2H, t, J=6.0Hz),6.54(1H, d, J=8.2Hz), 6.58(1H, d, J=8.2Hz), 7.23 (1H, t, J=8.2Hz),7.34(1H, s), 8.88 (1H, br s), 9.31(1H, br s). 2-56

464.45 351 A A (500 MHz, DMSO-d6): 1.65(1H, m), 1.78(1H, m), 1.93(2H, d,J=11.4Hz), 2.88(1H, m), 3.03(1H, m), 3.21(1H, m), 3.35(2H, dd, J=10,25Hz), 4.57 (2H, d, J=5.7Hz), 5.33(2H, m), 6.08 (1H, m), 6.56(2H, dd,J=8.3, 26Hz), 7.29(3H, M), 8.65(1H, br), 8.97 (1H, br). 2-57

360.85 325 A A (500 MHz, DMSO-d6): 1.67-1.96 (5H, m), 2.72-3.42(6H, m),3.79(3H, s), 6.55(1H, d, J=8.3Hz), 6.58(1H, d, J=8.3Hz), 7.22(1H, s),7.23(1H, t, J= 8.3Hz), 8.79(1H, br d), 9.13(1H, br d). 2-58

374.87 339 A A (500 MHz, DMSO-d6): 1.37(3H, t, J=6.9Hz), 1.68(1H, m),1.82(1H, m), 1.92-1.96(2H, m), 2.91(1H, m), 3.08 (1H, m), 3.23-3.32(2H,m), 3.39(1H, m), 4.06(2H, q, J=6.6Hz), 6.54(1H, d, J=8.2Hz), 6.58(1H, d,J=8.5Hz), 7.24(1H, dd, J=8.2, 8.5Hz), 7.40(1H, s), 7.46(1H, br s),8.81(1H, br s), 9.22(1H, br s). 2-59

402.93 367 A A (500 MHz, DMSO-d6): 0.92(6H, d, J=6.6Hz), 1.68(1H, m),1.80(1H, m), 1.91-1.94(2H, m), 2.04(1H, sep, J= 6.6Hz), 2.86(1H, m),3.07(1H, m), 3.27-3.36(3H, m), 3.76(2H, d, J=6.6 Hz), 6.56(1H, d,J=7.3Hz), 6.57(1H, d, J=8.2Hz), 7.23(1H, dd, J=7.3, 8.2 Hz), 7.28(1H,s), 7.56(1H, br s), 8.95 (1H, br s), 9.45(1H, br s). 2-60

416.96 381 A A (500 MHz, DMSO-d6): 0.84-0.86 (2H, m), 1.29-1.30(4H, m),1.69- 1.73(2H, m), 1.80-1.95(3H, m), 2.87 (1H, m), 3.10(1H, m),3.29-3.35(3H, m), 3.48(1H, m), 3.70(1H, m), 3.97 (2H, q, J=6.3Hz),6.59(2H, d, J=8.2 Hz), 7.25(1H, t, J=8.2Hz), 7.31(1H, s), 7.75(1H, brs), 9.07(1H, br s), 9.60(1H, br s). 2-61

457.02 421 A A (500 MHz, DMSO-d6): 0.85-0.93 (2H, m), 1.09-1.18(3H, m),1.32(1H, m), 1.60-1.64(7H, m), 1.71(1H, m), 1.84(1H, m), 1.90-1.95(2H,m), 2.87 (1H, m), 3.09(1H, m), 3.28(1H, m), 3.30-3.34(2H, m), 3.99(1H,dd, J= 6.6, 9.5Hz), 4.04(1H, dd, J=6.6, 9.5 Hz), 6.56 (1H, d, J=8.2Hz),6.60(1H, d, J=8.5Hz), 7.23(1H, dd, J= # 8.2, 8.5Hz), 7.30(1H, s),7.59(1H, br s), 8.99(1H, br s), 9.46(1H, br s). 2-62

414.94 379 A A (500 MHz, DMSO-d6): 1.63-1.78 (4H, m), 1.81-1.99(5H, m),2.02- 2.09(2H, m), 2.73(1H, m), 2.88(1H, m), 3.05(1H, m), 3.22-3.38(2H,m), 3.99(2H, d, J=6.6Hz), 6.53(1H, d, J= 8.5Hz), 6.59(1H, d, J=8.2Hz),7.22 (1H, t, 8.2Hz), 7.27(1H, s), 7.28(1H, br s), 8.76(1H, br s),9.11(1H, br s). 2-63

430.98 395 A A (500 MHz, DMSO-d6): 0.84(6H, d, J=6.6Hz), 1.17-1.25(2H,m), 1.52 (1H, sep, J=6.6Hz), 1.67-1.75(3H, m), 1.80(1H, m),1.86-1.96(2H, m), 2.87(1H, m), 3.07(1H, m), 3.24-3.39 (3H, m),3.87-4.13(2H, m), 6.54(1H, d, J=8.2Hz), 6.57(1H, d, J=8.2Hz), 7.22(1H,t, J=8.2Hz), 7.28(1H, s), 7.43, (1H, br s), 8.87 (1H, br s), # 9.27(1H,br s). 2-64

445.01 409 A B (500 MHz, DMSO-d6): 0.84(2H, d, J=6.9Hz), 0.85(2H, d,J=6.6Hz), 1.20-1.33(3H, m), 1.66-1.74(3H, m), 1.75-1.95(5H, m), 2.87(1H,m), 3.10 (1H, m), 3.22-3.47(4H, m), 3.90(1H, d, J=6.3Hz), 3.94-4.02(2H,m), 6.55 (1H, d, J=8.2Hz), 6.58(1H, d, J=8.2 Hz), 7.23(1H, t, J=8.2Hz),7.32(1H, s), 7.48(1H, br s), 8.98(1H, # m), 9.24 (1H, m). 2-65

400.91 365 A A (500 MHz, DMSO-d6): 0.33-0.37 (2H, m), 0.62-0.59(2H, m),1.30(1H, m), 1.67(1H, m), 1.82(1H, m), 1.91- 1.98(2H, m), 2.07(1H, m),2.88(1H, m), 3.11(1H, m), 3.26-3.32(2H, m), 3.85(2H, d, J=6.9Hz),6.53(1H, d, J= 8.5Hz), 6.54(1H, d, J=8.2Hz), 7.23 (1H, dd, J=8.2,8.5Hz), 7.37(1H, br s), 7.54(1H, s), 8.94(1H, m), # 9.43 (1H, m). 2-66

430.98 395 A A (500 MHz, DMSO-d6): 0.83(6H, d, J=6.6Hz), 1.03(2H, d,J=6.6Hz), 1.18-1.22(2H, m), 1.51-1.53(1H, m), 1.72-1.77(3H, m),1.93-1.95(4H, m), 2.84-2.85(1H, m), 3.06-3.22 (3H, m), 3.34-3.37(2H, m),3.99(1H, t, J=6.6Hz), 6.55-6.59(2H, m), 7.21- 7.26(2H, m), 9.03-9.12(1H,br s). 2-67

388.90 353 A A (500 MHz, DMSO-d6): 0.92(3H, t, J=7.25Hz), 1.77(2H, m),1.90-1.98 (2H, m), 3.06-3.13(3H, m), 3.38- 3.40(2H, m), 3.97(2H, t,J=6.3Hz), 6.57(2H, dd, J=10.0, 8.3Hz), 7.23- 7.27(2H, m), 9.06(2H, brs). 2-68

414.94 379 A A (500 MHz, DMSO-d6): 1.70-1.76 (2H, m), 1.79-1.92(4H, m),1.95- 1.98(2H, m), 2.00-2.07(2H, m), 2.77 (1H, m), 3.07-3.14(3H, m),3.39- 3.41(2H, m), 4.00(2H, d, J=6.6Hz), 6.56(1H, d, J=8.2Hz), 6.60(1H,d, J=8.2Hz), 7.19(1H, s), 7.24(1H, t, J=8.2Hz), 7.51(1H, br s), 9.03(2H,br s). 2-69

416.96 381 A A (500 MHz, DMSO-d6): 0.85(3H, dd, J=6.6, 7.2Hz),1.31-1.33(4H, m), 1.74-1.78(2H, m), 1.83-1.91(2H, m), 1.94-1.99(2H, m),3.08-3.12(3H, m), 3.37-3.39(2H, m), 4.00(2H, dd, J= 6.3, 6.6Hz),6.53(1H, d, J=8.2Hz), 6.58(1H, d, J=8.5Hz), 7.22(1H, dd, J=8.2, 8.5Hz),7.29(1H, s), 7.30(1H, br s), 8.75(1H, br s), 8.83(1H, br s). 2-70

457.02 421 A A (500 MHz, DMSO-d6): 0.86-0.93 (2H, m), 1.09-1.23(3H, m),1.34(1H, m), 1.59-1.67(7H, m), 1.86-1.99 (4H, m), 3.08-3.13(3H, m),3.36- 3.38(2H, m), 4.04(2H, dd, J=6.6, 6.9Hz), 6.54(1H, d, J=8.2Hz),6.60 (1H, d, J=8.5Hz), 7.23(1H, dd, J=8.2, 8.5Hz), 7.24(1H, s), 7.36(1H,br s), 8.91(2H, br s). 2-71

402.93 367 A A (500 MHz, DMSO-d6): 0.92(6H, d, J=6.6Hz), 1.83-1.92(2H,m), 1.94- 1.97(2H, m), 2.08(1H, dq, J=6.6Hz), 3.04-3.13(3H, m),3.38-3.41(2H, m), 3.78(2H, d, J=6.6Hz), 6.54(1H, d, J= 8.2Hz), 6.57(1H,d, J=8.5Hz), 7.22 (1H, s), 7.23(1H, dd, J=8.2, 8.5Hz), 7.38(1H, br s),8.86(1H, br s), 8.95 (1H, br s). 2-72

402.93 367 A (500 MHz, DMSO-d6): 0.89(3H, dd, J=7.3, 7.6Hz), 1.37(2H,dq, J=7.3, 7.6Hz), 1.75(2H, dd, J=6.6, 6.9Hz), 1.86-1.97(4H, m),3.06-3.13(3H, m), 3.37-3.40(2H, m), 4.02(2H, t, J= 6.6Hz), 6.54(1H, d,J=8.2Hz), 6.59 (1H, d, J=8.2Hz), 7.23(1H, t, J=8.2 Hz), 7.28(1H, s),7.47(1H, br s), 8.96 (2H, br s). 2-73

445.01 409 A A (300 MHz, DMSO-d6): 0.84(3H, t, J=6.8Hz), 1.22-1.27(8H,m), 1.73- 1.78(2H, m), 1.87-1.94(4H, m), 3.07-3.11(3H, m), 3.35-3.40(2H,m), 4.01(2H, t, J=6.4Hz), 6.54(1H, d, J= 8.3Hz), 6.58(1H, d, J=7.9Hz),7.23 (1H, dd, J=7.9, 8.3Hz), 7.28(1H, s), 7.33(1H, br s), 8.90(2H, brs). 2-74

386.88 351 A A (300 MHz, DMSO-d6): 1.78-1.98 (4H, m), 3.06-3.10(3H, m),3.38- 3.42(2H, m), 4.61(2H, d, J=5.3Hz), 5.29(1H, dd, J=1.5, 10.6Hz),5.36 (1H, dd, J=1.5, 17.3Hz), 6.11(1H, ddt, J=5.3, 10.6, 17.3Hz),6.55(1H, d, J=8.3Hz), 6.59(1H, d, J=7.9Hz), 7.23(1H, dd, J=7.9, 8.3Hz),7.27(1H, s), 7.33(1H, br s), 8.74(2H, br s). 2-75

374.87 339 A A (300 MHz, DMSO-d6): 1.40(3H, t, J=7.2Hz), 1.80-2.01(4H,m), 3.08- 3.10(3H, m), 3.39-3.43(2H, m), 4.08 (2H, q, J=6.8Hz), 6.53(1H,d, J=7.9 Hz), 6.57(1H, d, J=8.3Hz), 7.23(1H, dd, J=7.9, 8.3Hz), 7.34(1H,br s), 7.40(1H, s), 8.80(2H, br s). 2-76

430.98 395 A A (300 MHz, DMSO-d6): 0.36(3H, t, J=6.8Hz), 1.25-1.34(7H,m), 1.71- 1.78(2H, m), 1.82-1.91(2H, m), 1.94-1.98(2H, m), 3.05-3.37(4H,m), 4.01 (2H, t, J=6.4Hz), 6.52(1H, d, J= 8.3Hz), 6.58(1H, d, J=7.9Hz),7.20 (1H, br s), 7.22(1H, dd, J=7.9, 8.3 Hz), 7.30(1H, s), 8.61(1H, brs), 8.75 (1H, br s). 2-77

444.97 409 A A (500 MHz, DMSO-d6): 1.20(9H, s), 1.81-1.97(5H, m),2.98(1H, m), 3.26-3.31(2H, m), 3.42(1H, m), 5.20 (2H, s), 6.55(1H, d,J=8.2Hz), 6.61 (1H, d, J=8.2Hz), 7.25(1H, t, J=8.2 Hz), 7.36(2H, br s),8.13(1H, s), 8.81 (1H, br s), 9.27(1H, br s). 2-78

444.97 409 A A (300 MHz, DMSO-d6): 1.17(9H, s), 1.86-1.95(4H, m),3.07-3.10(3H, m), 3.35-3.40(2H, m), 5.22(2H, s), 6.44 (1H, d, J=8.3Hz),6.53(1H, d, J=8.3 Hz), 7.18(1H, t, J=8.3Hz), 7.21(1H, br s), 7.46(1H, brs), 8.77(1H, br s). 2-79

522.53 409 A A (500 MHz, DMSO-d6): 1.21-1.30 (2H, m), 1.43-1.51(3H, m),1.61(1H, m), 1.67-1.82(4H, m), 1.91-1.97(2H, m), 2.89(1H, m), 3.05(1H,m), 3.22 (1H, m), 3.35-3.43(3H, m), 3.91(1H, m), 3.96 (1H, m), 6.52(1H,d, J=8.5 Hz), 6.58(1H, d, J=8.5Hz), 7.22(1H, t, J=8.2Hz), 7.28(2H, brs), 7.38(1H, s), 8.67(1H, br s), 8.88(1H, br s). 2-80

508.50 395 A A (500 MHz, DMSO-d6): 1.61(1H, m), 1.66-1.81(4H, m),1.85(1H, m), 1.90-2.04(4H, m), 2.80(1H, m), 3.12 (1H, m), 3.23(1H, m),3.40(1H, m), 3.76(1H, m), 3.87(1H, m), 4.03(1H, dd, J=2.8, 3.1Hz),4.27(1H, m), 6.52 (1H, d, J=8.2Hz), 6.55(1H, d, J=8.5 Hz), 7.24(1H, dd,J=8.2, 8.5Hz), 7.29 (2H, br s), 7.61(1H, s), 8.62(1H, br # s), 8.92(1H,br s). 2-81

430.94 395 A A (500 MHz, DMSO-d6): 1.61(1H, m), 1.71(1H, m),1.76-1.86(3H, m), 1.87-2.03(3H, m), 2.78(1H, m), 3.13 (1H, m),3.25-3.35(3H, m), 3.75- 3.80(2H, m, 3.88(1H, m), 4.03(1H, m), 4.27(1H,m), 7.51(1H, d, J=8.2 Hz), 7.56(1H, d, J=8.2Hz), 7.24(1H, t, J=8.2Hz),7.53(1H, br s), 7.58(1H, s), 8.93(1H, m), 9.53(1H, m). 2-82

430.94 395 A A (300 MHz, DMSO-d6): 1.59(1H, m), 1.76-1.97(7H, m),3.07-3.10(3H, m), 3.37-3.41(2H, m), 3.70(2H, dd, J= 6.8, 14.7Hz),3.92(1H, dd, J=6.8, 7.2 Hz), 4.01(1H, dd, J=6.8, 7.2Hz), 4.22 (1H, m),6.54(1H, d, J=8.3Hz), 6.58 (1H, d, J=8.3Hz), 7.20(1H, br s), 7.22(1H, t,J=8.3Hz), 7.34(1H, s), 8.74(1H, br s), 8.92(1H, br s). 2-83

494.47 422 B B (500 MHz, DMSO-d6): 1.26-1.38 (1H, m), 1.65-1.93(10H, m),2.84- 3.39(8H, m), 4.36-4.43(3H, m), 6.61 (2H, d, J=8.2Hz), 7.05(1H, s),7.22- 7.26(3H, m), 8.78-8.85(1H, m), 9.38 (1H, br d, J=10.1Hz),10.56(1H, br s). 2-84

496.44 424 A B (500 MHz, DMSO-d6): 1.71-1.93 (5H, m), 2.95-3.47(12H, m),3.75 (1H, br s), 4.36-4.44(2H, br s), 6.61 (2H, d, J=8.2Hz), 7.01(1H,s), 7.16 (2H, br s), 7.23(1H, t, J=8.4Hz), 8.76-8.85(1H, m), 9.29(1H, brd, J=10.4Hz), 11.34(1H, br s). 2-85

417.94 382 B B (500 MHz, DMSO-d6): 1.68-1.83 (2H, m), 1.92(2H, m),2.73(6H, d, J= 4.4Hz), 2.97(1H, m), 3.17-3.19(3H, m), 3.22-3.39(3H, m),4.36(2H, br), 6.61(1H, d, J=8.5Hz), 6.63(1H, d, J=8.2Hz), 7.09(1H, s),7.21(1H, br s), 7.25(1H, dd, J=8.2, 8.5Hz), 8.80 (1H, br), 9.37(1H, brs), 10.6(1H, br s). 2-86

472.04 436 C B (500 MHz, DMSO-d6): 1.53-1.54 (2H, m), 1.62-1.64(2H, m),1.67- 1.84(6H, m), 1.91-1.92(2H, m), 2.97 (1H, m), 3.05-3.10(2H, m),3.17- 3.25(2H, m), 3.29-3.45(6H, m), 4.39 (2H, br), 6.60(1H, d,J=8.2Hz), 6.61 (1H, d, J=8.2Hz), 7.05(1H, s), 7.18 (1H, br s), 7.24(1H,t, J=8.2Hz), 8.77 (1H, br s), 9.30(1H, br s), # 10.6 (1H, br s). 2-87

443.98 408 C B (500 MHz, DMSO-d6): 1.72-1.85 (5H, m), 1.90-1.93(3H, m),2.91- 3.17(3H, m), 3.21-3.39(4H, m), 3.43-3.44(2H, m), 3.50-3.51(2H, m),4.36(2H, br), 6.62(1H, d, J=8.5Hz), 6.65(1H, d, J=8.2Hz), 7.15(1H, s),7.26(1H, dd, J=8.2, 8.5Hz), 7.53(1H, br s), 9.00(1H, br s), 9.64(1H, brs), 11.2(1H, br s).

Example 3-1

(1) With the use of the starting compound 1G, 2B and other materials,2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]4-(3-piperidinyl)nicotinonitrilehydrochloride was prepared in a similar manner as described in Example2-3.

(2) To a stirred solution of2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-(3-piperidinyl)nicotinonitrilehydrochloride (200.0 mg, 0.50 mmol) in MeOH were added formaldehyde (0.5mL) and sodium cyanoborohydride (40.8 mg, 0.65 mmol) and stirred for 2hrs at room temperature. The reaction was quenched by an addition ofwater, extracted with ethyl acetate, dried over MgSO₄, filtered andevaporated. The residue was triturated with hexane and dried. The crudeproduct was purified by preparative silica gel TLC (5% MeOH indichloromethane) to give2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-(1-methyl-3-piperidinyl)nicotinonitrileas a yellow solid. (28.5 mg, yield 15%)

Molecular weight: 378.48; Mass spectometry: 379 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)A; ¹H-NMR (300 MHz,CDCl3): 0.37-0.42 (2H, m), 0.67-0.73 (2H, m), 0.82-0.88 (1H, m),1.27-1.35 (1H, m), 1.76-2.09 (4H, m), 2.31 (3H, s), 2.91-3.00 (2H, m),3.14-3.23 (1H, m), 3.86 (2H, pent, J=8.8 Hz), 5.15 (2H, br s), 6.39 (1H,d, J=8.3 Hz), 6.62 (1H, d, J=8.3 Hz), 7.20 (1H, t, J=8.3 Hz), 7.95 (1H,s).

Example 3-2

To a stirred solution of2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-(3-piperidinyl)nicotinonitrilehydrochloride (200.0 mg, 0.50 mmol), which was obtained in the step (1)of example 3-1, in MeOH was added benzaldehyde (0.25 mL, 2.49 mmol))followed by sodium cyanoborohydride (40.8 mg, 0.65 mmol), and thestirring was continued for 2 hrs at room temperature. The reaction wasquenched by an addition of water, extracted with ethyl acetate, driedover MgSO₄, filtered and evaporated. The residue was triturated withhexane and dried. The crude product was purified by preparative silicagel TLC (5% MeOH in dichloromethane) to give2-amino-4-(1-benzyl-3-piperidinyl)-6-[2-(cyclopropylmethoxy)-6-hydroxy-phenyl]nicotinonitrileas a yellow solid. (35.7 mg, yield 16%)

Molecular weight: 454.58; Mass spectrometry: 455 (M+H)⁺; In vitroactivity grade: C; Cellular activity grade: (A549)C; ¹H-NMR (300 MHz,CDCl3): 0.35-0.37 (2H, m), 0.62-0.64 (2H, m), 0.83-1.29 (2H, m),1.75-2.21 (5H, m), 2.93-3.24 (3H, m), 3.50-3.61 (2H, m), 3.78-3.88 (2H,m), 5.12 (2H, br s), 6.39 (1H, d, J=8.3 Hz), 6.60 (1H, d, J=8.3 Hz),7.16-7.32 (6H, m), 7.93 (1H, s), 13.33 (1H, br s).

Examples 3-3 to 3-8

According to the similar synthetic procedure of Examples 3-1 to 3-2,compounds shown in Table 3 were prepared.

TABLE 3 Mol In Ex. No Structure weight Mass vitro A549 NMR 3-03

344.85 309 A A (300 MHz, DMSO-d6): 1.70-1.96 (4H, m), 2.73(3H, d,J=1.8Hz), 2.93-2.98(1H, m), 3.35-3.49 (4H, m), 6.90-6.95(2H, m), 7.33-7.46(3H, m), 8.02(1H, dd, J= 8.5, 1.3Hz), 10.62(1H, br s). 3-04

372.90 337 A B (300 MHz, DMSO-d6): 0.92(3H, t, J=7.4Hz), 1.71-1.76(3H,m), 1.81-1.98(3H, m), 2.92-3.04 (3H, m), 6.90-6.95(2H, m), 7.34-7.38(2H, m), 7.45(1H, br s), 8.01(1H, d, J=7.4Hz), 9.80(1H, br s). 3-05

420.95 385 A A (300 MHz, DMSO-d6): 1.81-1.96 (4H, m), 2.72-2.73(1H, m),3.31- 3.50(4H, m), 4.26-4.35(2H, m), 6.89-6.95(2H, m), 7.34- 7.51(6H,m), 7.59-7.62(2H, m), 8.00-8.02(1H, m), 10.57(1H, br s). 3-06

434.97 399 B B (300 MHz, DMSO-d6): 1.78-2.01 (5H, m), 3.04-3.15(4H, m),3.30- 3.67(2H, m), 6.90-6.95(2H, m), 7.24-7.49(11H, m), 8.03 (1H, d,J=7.5Hz), 10.58(1H, br s). 3-07

430.51 431 A B (500 MHz, DMSO-d6): 1.39(1H, dd, J=3.5, 11.7Hz), 1.59(1H,m), 1.73(1H, d, J=12.9Hz), 1.88(1H, d, J=11.7Hz), 1.95 (1H, t,J=10.4Hz), 2.02(1H, t, J= 11.0Hz), 2.84(1H, d, J=11.0 Hz), 2.89(1H, d,J=10.4Hz), 2.97(1H, t, J=11.0Hz), 3.45 (2H, m), 3.72(3H, s), 6.36(2H, d,J=8.2Hz), 6.86(2H, d, J=8.5 Hz), # 7.06(1H, t, J=8.2Hz), 7.16 (2H, br),7.21(1H, d, J=8.5Hz), 7.78(1H, s), 11.91(2H, s). 3-08

484.60 485 C C (300 MHz, CDCl3): 0.33-0.38 (2H, m), 0.60-0.66(2H, m),1.24- 2.21(8H, m), 2.94-3.25(2H, m), 3.52(2H, br s), 3.79-3.84 (5H, m),5.16(2H, br s), 6.39(1H, d, J=8.3Hz), 6.60(1H, d, J=8.3 Hz), 6.84(2H, d,J=8.7Hz), 7.16- 7.26(3H, m), 7.93(1H, s).

Example 4-1

(1) A mixture of copper (II) bromide (1.106 g, 4.953 mmol) andtert-butyl nitrite (0.736 mL, 6.191 mmol) in acetonitrile (30 mL) wasstirred at 65° C. for 15 min. A solution of tert-butyl3-{2-amino-6-[2-(benzyloxy)phenyl]-3-cyano-4-pyridinyl}-1-piperidinecarboxylate(2.000 g, 4.127 mmol), which was obtained in the step (1) of Example1-1, in acetonitrile (20 mL) was added dropwise to the mixture. Themixture was stirred at 65° C. for 2 hrs. After cooled to roomtemperature, the reaction mixture was diluted with ethyl acetate. Theseparated organic phase was washed with an aqueous 1N HCl solution andbrine, dried over Na₂SO₄, filtered, and concentrated. The residue wasrecrystallized from diethyl ether to give tert-butyl3-{2-bromo-6-[2-(benzyloxy)phenyl]-3-cyano-4-pyridinyl}-1-piperidinecarboxylateas a white solid (1.010 g, yield; 45%).

(2) To a stirred solution of tert-butyl3-{6-[2-(benzyloxy)phenyl]-2-bromo-3-cyano-4-pyridinyl}-1-piperidinecarboxylate(0.300 g, 0.547 mmol) in DMSO were added triethylamine (0.229 mL, 1.341mmol) and benzylamine (0.147 g, 1.367 mmol). The mixture was stirred at70° C. for 12 hrs. After cooled to room temperature, the reactionmixture was quenched with water and extracted with ethyl acetate. Theseparated organic phase was washed with brine, dried over Na₂SO₄,filtrated, and concentrated. The residue was purified by silica gelcolumn chromatography (hexane/ethyl acetate=4/1-2/1) to give tert-butyl3-{6-[2-(benzyloxy)phenyl]-2-benzylamino-3-cyano-4-pyridinyl}-1-piperidinecarboxylateas a yellow amorphous. (0.290 g, yield; 92%)

(3) Then benzyl moiety was removed in a same manner as described in step(2) of Example 1-1. The residue was suspended in ethanol and filtratedto give tert-butyl3-[2-(benzylamino)-3-cyano-6-(2-hydroxyphenyl)-4-pyridinyl]-1-piperidinecarboxylateas a white solid. (0.150 g, yield; 61%)

(4) tert-butyl3-[2-(benzylamino)-3-cyano-6-(2-hydroxyphenyl)-4-pyridinyl]-1-piperidinecarboxylate(0.145 g, 0.299 mmol) was treated under acidic conditions in a similarmanner as described in step (3) in Example 1-1. The resulting solid wascollected by filtration, and dried under reduced pressure to give2-(benzylamino)-6-(2-hydroxyphenyl)-4-(3-piperidinyl)nicotinonitrilehydrochloride. (0.075 g, yield; 60%).

Molecular weight: 420.95; Mass spectrometry: 385 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-B/Jurkat-B; ¹H-NMR(300 MHz, DMSO-d6): 1.80-1.92 (4H, m), 2.90-2.93 (1H, m), 3.31-3.40 (4H,m), 4.59 (2H, d, J=5.6 Hz), 6.84-6.92 (2H, m), 7.19-7.31 (5H, m), 7.47(1H, s), 8.00 (1H, d, J=6.8 Hz), 8.11-8.15 (1H, m), 8.91 (1H, br s),9.52 (1H, br s), 12.59 (1H, br s), 12.90 (1H, br s).

Example 4-2

To a stirred solution of tert-butyl3-{6-[2-(benzyloxy)phenyl]-2-bromo-3-cyano-4-pyridinyl}-1-piperidinecarboxylate(0.100 g, 0.182 mmol), which was obtained in the steps (1) of Example4-1, in DMSO (2 mL) were added propylamine (0.108 g, 1.823 mmol) andtriethylamine (0.038 mL, 0.273 mmol). The mixture was stirred at 40° C.for 12 hrs. The reaction was quenched with water and the resultingreaction mixture was extracted with ethyl acetate. The organic phase wasdried over Na₂SO₄, filtered, and concentrated. The concentrates waspurified by silica gel column (hexane/ethyl acetate=4/1) to givetert-butyl3-{6-[2-(benzyloxy)phenyl]-3-cyano-2-propylamino-4-pyridinyl}-1-piperidinecarboxylateas a colorless oil. (0.104 g, yield; quant.)

Then benzyl moiety was removed in a same manner as described in the step(2) in Example 1-1. The residue was washed with diethyl ether to givethe desired product as a white solid. (0.112 g, yield; quant.)

Then tert-butyl3-[3-cyano-6-(2-hydroxyphenyl)-2-(propylamino)-4-pyridinyl]-1-piperidinecarboxylate(0.110 g, 0.252 mmol) was treated under acidic conditions in a samemanner as described in the step (3) in Example 1-1. The resulting solidwas collected with filtration, and dried under reduced pressure to give6-(2-hydroxyphenyl)-4-(3-piperidinyl)-2-(propylamino)nicotinonitrilehydrochloride. (0.062 g, yield; 66%)

Molecular weight: 372.90; Mass spectrometry: 337 (M+H)⁺; In vitroactivity grade: C; Cellular activity grade: (A549)-C; ¹H-NMR (300 MHz,DMSO-d6): 0.92 (3H, t, J=7.4 Hz), 1.62 (2H, m), 1.80-1.95 (4H, m),2.89-2.93 (1H, m), 3.33-3.46 (6H, m), 6.91-6.97 (2H, m), 7.34-7.40 (1H,m), 7.48 (1H, s), 7.57 (1H, br s), 8.08-8.17 (1H, m), 8.97-9.07 (1H, m),9.67-9.71 (1H, m).

Example 4-3

In a similar manner as that of Example 4-2,6-(2-hydroxyphenyl)-2-(methylamino)-4-(3-piperidinyl)nicotinonitrilehydrochloride was prepared.

Molecular weight: 344.85; Mass spectrometry: 309 (M+H)⁺; In vitroactivity grade: C; Cellular activity grade: (A549)-C; ¹H-NMR (300 MHz,DMSO-d6): 1.83-1.96 (4H, m), 3.31 (4H, s), 3.30-3.56 (4H, m), 6.90-6.96(2H, m), 7.34-7.37 (1H, m), 7.39 (1H, s), 7.55 (1H, br s), 8.10 (1H, d,J=7.9 Hz), 8.87-8.90 (1H, m), 9.52-9.54 (1H, m), 13.80 (1H, br s).

Example 4-4

In a similar manner as that of Example 4-2,2-anilino-6-(2-hydroxyphenyl)-4-(3-piperidinyl)nicotinonitrilehydrocloride was prepared.

Molecular weight: 406.92; Mass spectrometry: 371 (M+H)⁺; In vitroactivity grade: D; Cellular activity grade: (A549)-B; ¹H-NMR (300 MHz,DMSO-d6): 1.87-2.00 (4H, m), 2.91-2.95 (1H, m), 3.34 (1H, m), 6.79 (1H,d, J=8.3 Hz), 6.91 (1H, t, J=7.2 Hz), 7.18-7.44 (1H, brs), 7.66 (1H, s),8.06 (1H, d, J=6.8 Hz), 8.84 (1H, m), 9.43 (2H, m), 12.42 (1H, br s).

Example 4-5

In a similar manner as that of Example 4-2,6-(2-hydroxyphenyl)-2-(1-piperazinyl)-4-(3-piperidinyl)nicotinonitriledihydrochloride was prepared.

Molecular weight: 436.39; Mass spectrometry: 364 (M+H)⁺; In vitroactivity grade: D¹H-NMR (500 MHz, DMSO-d6): 1.81-1.83 (2H, m), 1.90-1.95(2H, m), 2.93-2.95 (1H, m), 3.76-3.78 (4H, m), 6.98 (1H, t, J=7.3 Hz),7.37-7.40 (1H, m), 7.84 (1H, s), 8.03-8.05 (1H, m), 8.78-8.81 (1H, m),9.14 (1H, br s), 9.24-9.27 (1H, m), 12.10 (1H, s).

Example 5-1

(1) A mixture of tert-butyl3-{6-[2-(benzyloxy)phenyl]-2-bromo-3-cyano-4-pyridinyl}-1-piperidinecarboxylate(0.300 g, 0.547 mmol), which was obtained in the step (1) of Example4-1, hydrazine monohydrate (3 mL) and 1,4-dioxane (1 mL) was stirred at100° C. for 1.5 hrs. The reaction was quenched with water and extractedwith ethyl acetate. The organic phase was washed with brine and driedover Na₂SO₄, filtered, and concentrated. The residue was recrystallizedfrom ethanol to give tert-butyl{3-[3-amino-6-(2-benzyloxy-phenyl)-1H-pyrazolo[3,4-b]pyridin-4-yl]-1-piperidinecarboxylateas a yellow solid. (0.210 g, yield; 72%).

(2) Then the benzyl moiety was removed in a same manner as described inthe step (2) of Example 1-1. The residue was purified by silica gelcolumn (hexane/ethyl acetate=2/1-1/1) to give tert-butyl3-[3-amino-6-(2-hydroxyphenyl)-1H-pyrazolo[3,4-b]pyridin-4-yl]-1-piperidinecarboxylateas an orange solid. (0.080 g, yield; 49%).

To a stirred solution tert-butyl3-[3-amino-6-(2-hydroxyphenyl)-1H-pyrazolo[3,4-b]pyridin-4-yl]-1-piperidinecarboxylate(0.075 g, 0.183 mmol) in 1,4-dioxane (3 mL) was added 4N HCl in1,4-dioxane (3 mL). The mixture was stirred at room temperature for 12hrs. The resulting solid was collected with filtration to give2-[3-amino-4-(3-piperidinyl)-1H-pyrazolo[3,4-b]pyridin-6-yl]phenolhydrochloride. (0.062 g, yield; 98%)

Molecular weight: 345.83; Mass spectrometry: 310 (M+H)⁺; In vitroactivity grade: C; Cellular activity grade: A549-B; ¹H-NMR (300 MHz,DMSO-d6): 1.97-2.27 (4H, m), 2.95-3.11 (1H, m, 3.12-3.23 (1H, m),3.30-3.39 (3H, m), 6.94-6.99 (2H, m), 7.32-7.38 (1H, m, 7.73 (1H, s),8.12 (1H, d, J=7.2 Hz).

Example 5-2

To a cooled (0° C.), stirred solution of tert-butyl3-{3-amino-6-[2-(benzyloxy)phenyl]-1H-pyrazolo[3,4-b]pyridin-4-yl}-1-piperidinecarboxylate(0.200 g, 0.400 mmol) obtained in the step (1) of Example 5-1 in THF (3mL) were added pyridine (1 mL) and acetyl chloride (0.035 g, 0.440mmol). The mixture was stirred at 0° C. for 1 hr. The reaction wasquenched with water and extracted with ethyl acetate. The organic phasewas washed with saturated brine, dried over Na₂SO₄, filtered, andconcentrated. The residue was recrystallized from diisopropyl ether togive a white solid. (0.180 g, yield; 83%).

Then the benzyl moiety was removed in a same manner as described in thestep (2) of Example 1-1. The residue was suspended in ethanol andfiltrated to give the desired product as a white solid. (0.120 g, yield;82%).

tert-Butyl3-[3-(acetylamino)-6-(2-hydroxyphenyl)-1H-pyrazolo[3,4-b]pyridin-4-yl]-1-piperidinecarboxylate(0.115 g, 0.260 mmol) was treated under acidic conditions in a samemanner as described in the step (3) of Example 1-1 to giveN-[6-(2-hydroxyphenyl)-4-(3-piperidinyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]acetamidehydrochloride. (0.097 g, yield; 96%)

Molecular weight: 387.87; Mass spectrometry: 352 (M+H)⁺; In vitroactivity grade: D; Cellar activity grade: A549-C; ¹H-NMR (500 MHz,DMSO-d6): 1.80-1.91 (4H, m), 1.95 (3H, s), 2.82-2.92 (1H, m), 3.34-3.44(4H, m), 6.92-6.95 (2H, m), 7.36-7.39 (1H, m), 7.66 (1H, s), 8.12 (1H,d, J=8.2 Hz), 8.83-8.85 (1H, m), 9.48-9.49 (1H, m), 9.56 (1H, s) 10.21(1H, s), 13.59 (1H, br s).

Example 6-1

To a cooled (0° C.), stirred solution of tert-butyl3-{2-amino-6-[2-(benzyloxy)phenyl]-3-cyano-4-pyridinyl}-1-piperidinecarboxylate,which was obtained in the step (1) of Example 1-1, (0.500 g, 1.032 mmol)in pyridine (10 mL) was added acetyl chloride (0.477 mL, 6.70 mmol). Themixture was stirred at 0° C. to room temperature for 3 hrs, and thestirring was continued at room temperature for 5 hrs. The reactionmixture was quenched with water, and extracted with ethyl acetate. Theorganic phase was dried over Na₂SO₄, filtered, and concentrated. Theresidue was purified by column chromatography on silica gel(hexane/ethyl acetate=9/1-4/1-2/1) to give a mixture of the desiredproduct, tert-butyl3-{2-(acetylamino)-6-[2-(benzyloxy)phenyl]-3-cyano-4-pyridinyl}-1-piperidinecarboxylate,and the corresponding diacylated compound. The mixture obtained wasdissolved in THF (5 mL), treated with an aqueous 1N NH₃ solution (1 mL),and stirred at room temperature for 1 hr. The reaction mixture wasextracted with ethyl acetate. The organic phase was washed with brine,dried over Na₂SO₄, filtered and concentrated to give tert-butyl3-{2-(acetylamino)-6-[2-(benzyloxy)phenyl]-3-cyano-4-pyridinyl}-1-piperidinecarboxylateas a white amorphous. (0.432 g, yield; 80%).

Then the benzyl moiety was removed in a same manner as described in thestep (2) of Example 1-1. The residue was recrystallized from ethanol togive the desired product as a yellow solid. (0.110 g, yield; 31%).

tert-Butyl3-[2-(acetylamino)-3-cyano-6-(2-hydroxyphenyl)-4-pyridinyl]-1-piperidinecarboxylate(0.100 g, 0.229 mmol) was treated under acidic conditions in a similarmanner as described in the step (3) of Example 1-1. The resultingprecipitate was collected by filtration, washed with 1,4-dioxane, anddried under reduced pressure to giveN-[3-cyano-6-(2-hydroxyphenyl)-4-(3-piperidinyl)-2-pyridinyl]acetamidehydrochloride. (0.101 g, yield; quant.)

Molecular weight: 372.86; Mass spectrometry: 337 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-B; ¹H-NMR ‘(500 MHz,DMSO-d6): 1.86-1.97 (4H, m), 2.19 (3H, s), 2.91-2.94 (1H, m), 3.33-3.47(4H, m), 6.96-6.99 (2H, m), 7.38-7.41 (1H, m), 8.11-8.13 (2H, m),8.94-8.96 (1H, m), 9.62-9.64 (1H, m), 10.97 (1H, s).

Example 6-2

With the use of the starting compound 1G and the 4-formyl piperidine-1carboxylic acid tert-butyl ester and other materials,N-[3-cyano-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-(4-piperidinyl)-2-pyridinyl]acetamidehydrochloride was prepared in the similar manner as that of Example 6-1.

Molecular weight: 442.95; Mass spectrometry: 407 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 0.28-0.31 (2H, m), 0.55-0.58 (2H, m), 1.32 (1H, m), 1.98-2.01(2H, m), 2.16 (3H, s), 3.09-3.13 (2H, m), 3.24-3.28 (1H, m), 3.40-3.42(2H, m), 3.87 (2H, d, J=6.9 Hz), 6.57 (2H, dd, J=8.5, 2.8 Hz), 7.24 (1H,t, J=8.5 Hz), 7.92 (1H, s), 9.06-9.15 (2H, m), 10.93 (1H, s).

Example 7-1

(1) To a stirred suspension of tert-butyl3-{2-amino-6-[2-(benzyloxy)phenyl]-3-cyano-4-pyridinyl}-1-piperidinecarboxylate(0.300 g, 0.619 mmol), which was obtained in the step (1) of Example1-1, in ethanol (10 mL) was added a solution of potassium hydroxide(0.695 g, 12.381 mmol) in ethanol (20 mL). The reaction mixture wasstirred at 70° C. for 60 hrs. After cooled to room temperature, thereaction mixture was poured into water and the resulting solid wasfiltrated and dried under reduced pressure to give tert-butyl3-{2-amino-6-[2-(benzyloxy)phenyl]-3-carbamoyl-4-pyridinyl}-1-piperidinecarboxylatas a white amorphous. (0.226 g, yield; 73%).

(2) Then benzyl moiety was removed in a same manner as described in thestep (2) of Example 1-1. The residue was suspended in ethanol andfiltrated to give tert-butyl3-[2-amino-3-(carbamoyl)-6-(2-hydroxyphenyl)-4-pyridinyl]-1-piperidinecarboxylateas a white solid. (0.045 g, yield; 46%).

(3) tert-butyl3-[2-amino-3-(carbamoyl)-6-(2-hydroxyphenyl)-4-pyridinyl]-1-piperidinecarboxylate(0.040 g, 0.097 mmol) was treated under acidic conditions in a similarmanner as described in the step (3) of Example 1-1 to give2-amino-6-(2-hydroxyphenyl)-4-(3-piperidinyl)nicotinamide hydrochlorideas a yellow solid. (0.008 g, yield; 24%)

Molecular weight: 348.84; Mass spectrometry: 313 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-B; ¹H-NMR (300 MHz,DMSO-d6): 1.68-1.92 (4H, m), 2.90-2.93 (1H, m), 3.17-3.34 (4H, m),6.89-6.97 (2H, m), 7.29-7.34 (2H, m), 7.80-7.86 (2H, m), 8.09 (1H, brs),8.80-8.82 (1H, m), 9.34-9.37 (1H, m).

Example 7-2

To a cooled (0° C.), stirred solution of tert-butyl3-{2-amino-6-[2-(benzyloxy)phenyl]-3-carbamoyl-4-pyridinyl}-1-piperidinecarboxylate(0.400 g, 0.800 mmol), which was obtained in the step (1) of Example7-1, in THF (10 mL) were added triethylamine (0.50 mL, 3.58 mmol)followed by triphosgene (0.354 g, 1.19 mmol). The reaction mixture wasstirred at room temperature for 12 hrs. The reaction was quenched withwater and extracted with ethyl acetate. The organic phase was washedwith brine, dried over Na₂SO₄, filtered, and concentrated. The residuewas purified by silica gel column chromatography (hexane/ethylacetate=2/1-1/1) and recrystallized from ethanol to give tert-butyl3-{7-[2-(benzyloxy)phenyl]-2,4-dioxo-1,2,3,4-tetrahydropyrido[2,3-d]pyrimidin-5-yl}-1-piperidinecarboxylateas a white solid. (0.406 g, yield; 97%).

Then the benzyl moiety was removed in a same manner as described in thestep (2) of Example 1-1. The residue was washed with ethanol to givetert-butyl3-[7-(2-hydroxyphenyl)-2,4-dioxo-1,2,3,4-tetrahydropyrido[2,3-d]pyrimidin-5-yl]-1-piperidinecarboxylateas a white solid. (0.036 g, yield; 29%).

tert-butyl3-[7-(2-hydroxyphenyl)-2,4-dioxo-1,2,3,4-tetrahydropyrido[2,3-d]pyrimidin-5-yl]-1-piperidinecarboxylate(0.036 g, 0.80 mmol) in dioxane (3 mL) was treated under acidicconditions in a similar manner as described in the step (3) of Example1-1 to give7-(2-hydroxyphenyl)-5-(3-piperidinyl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dionehydrochloride as a yellow solid. (0.016 g, yield; 52%)

Molecular weight: 374.83; Mass spectrometry: 339 (M+H)⁺; In activitygrade: A; Cellular activity grade: (A549)-C; ¹H-NMR (500 MHz, DMSO-d6):1.78-1.83 (2H, m), 1.94-1.95 (2H, m), 2.90-2.92 (1H, m), 4.68-4.69 (1H,m), 6.97-6.99 (2H, m), 7.39-7.42 (1H, m), 7.85 (1H, s), 8.16 (1H, dd,J=8.4, 1.4 Hz), 8.75 (1H, br s), 9.43 (1H, m), 11.45 (1H, d, J=1.4 Hz),12.05 (1H, d, J=1.4 Hz), 12.44 (1H, br s).

Example 7-3

To a stirred solution of tert-butyl3-{2-amino-6-[2-(benzyloxy)phenyl]-3-carbamoyl-4-pyridinyl}-1-piperidinecarboxylate(0.100 g, 0.199 mmol), which was obtained in the step (1) of Example7-1, in phenyl ether (1 mL) was added phenyl isocyanate (0.047 g, 0.398mmol). The mixture was stirred at 140° C. for 12 hrs. After cooled toroom temperature, the resulting solid (urea) was removed by filtration.The filtrate was concentrated under reduced pressure. The residue waspurified by silica gel column chromatography to give tert-butyl3-[7-(2-benzyloxyphenyl)-2,4-dioxo-3-phenyl-1,2,3,4-tetrahydropyrido[2,3-d]pyrimidin-5-yl]-1-piperidinecarboxylateas a white amorphous. (0.090 g, yield; 78%).

Then benzyl moiety was removed in a same manner as described in the step(2) of Example 1-1. The residue was washed with ethanol to give thedesired product as a white solid. (0.060 g, yield; 83%).

tert-butyl3-[7-(2-hydroxyphenyl)-2,4-dioxo-3-phenyl-1,2,3,4-tetrahydropyrido[2,3-d]pyrimidin-5-yl]-1-piperidinecarboxylate(0.050 g, 0.100 mmol) was treated under acidic conditions in a similarmanner as described in the step (3) of Example 1-1 to give7-(2-hydroxyphenyl)-3-phenyl-5-(3-piperidinyl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dionehydrochloride as a yellow solid. (0.016 mg, yield; 37%)

Molecular weight: 450.93; Mass spectrometry: 415 (M+H)⁺; In vitroactivity grade: B; Cellular activity grade: (A549)-C; ¹H-NMR (300 MHz,DMSO-d6): 1.71-1.96 (4H, m), 2.71-2.91 (1H, m), 3.27-3.53 (3H, m), 4.56(1H, m), 6.98-7.03 (2H, m), 7.34-7.53 (6H, m), 7.89 (1H, s), 8.18-8.21(1H, m), 8.69-8.70 (1H, m), 9.24-9.25 (1H, m), 12.47 (1H, s).

Example 8-1

(1) A solution of 1-{2-[(4-methoxybenzyl)oxy]phenyl}ethanone (14.2 g,55.4 mmol)(starting compound 1D), benzyl3-formyl-1-piperidinecarboxylate (13.7 g, 55.4 mmol)(starting compound2C), tert-butyl cyanoacetate (7.8 g, 55.4 mmol), and ammonium acetate(9.8 g, 166.1 mmol) in 1,2-dimethoxyethane (60 mL) was stirred underreflux for 3.5 hrs. After cooled to room temperature, the mixture waspartitioned between ethyl acetate and water. The separated organic phasewas washed with water and brine, dried over MgSO₄, filtered, andconcentrated under reduced pressure. The residue was purified by columnchromatography on silica gel (ethyl acetate:hexane, 1:2) to givetert-butyl2-amino-4-{1-[(benzyloxy)carbonyl]-3-piperidinyl}-6-{2-[(4-methoxybenzyl)oxy]phenyl}nicotinateas a pale yellow oil (4.89 g, yield; 14%).

(2) To a solution of tert-butyl2-amino-4-{1-[(benzyloxy)carbonyl]-3-piperidinyl}-6-{2-[(4-methoxybenzyl)oxy]phenyl}nicotinate(0.95 g, 1.67 mmol) in CH₂Cl₂ (10 mL) was added trifluoroacetic acid (10mL), and the stirring was continued at room temperature overnight. Themixture was concentrated under reduced pressure. The residue was dilutedwith toluene, then concentrated under reduced pressure to remove excessof trifluoroacetic acid by azeotropic distillation to give2-amino-4-{1-[(benzyloxy)carbonyl]-3-piperidinyl}-6-(2-hydroxyphenyl)nicotinicacid (1.1 g, yield; quant.).

(3) To a solution of2-amino-4-{1-[(benzyloxy)carbonyl]-3-piperidinyl}-6-(2-hydroxyphenyl)nicotinicacid (1.1 g, 2.5 mmol) in THF (10 mL) and MeOH (5 mL) was added dropwisetrimethylsilyldiazomatane (4.0 mL) at room temperature. The reactionmixture was stirred at room temperature for 1.5 hrs, and the reactionwas quenched by acetic acid. The mixture was partitioned between ethylacetate and water. The separated organic phase was washed with brine,dried over MgSO₄, filtered, and concentrated under reduced pressure. Theresidue was purified by column chromatography on silica gel(hexane:ethyl acetate, 3:1) followed by recrystallization from a mixtureof CH₂Cl₂ and hexane to give methyl2-amino-4-{1-[(benzyloxy)carbonyl]-3-piperidinyl}-6-(2-hydroxyphenyl)nicotinateas a yellow solid (0.39 g, yield; 34%).

(4) To a solution of methyl2-amino-4-{1-[(benzyloxy)carbonyl]-3-piperidinyl}-6-(2-hydroxyphenyl)nicotinate(60 mg, 0.130 mmol) in methanol (2.0 mL) and THF (1.0 mL) was added 10%Pd—C (200 mg). The mixture was stirred at room temperature under ahydrogen atmosphere (1 atm) for 6.5 hrs. The mixture was filtrated onCelite®, and washed with MEOH and THF successively. The filtrate wasconcentrated under reduced pressure. The residue was purified bypreparative TLC (hexane:ethyl acetate, 2:1) to give6-amino-4-(2-hydroxy-phenyl)-5,9-diazatricyclo[7.3.1.0^(2,7)]trideca-2(7),3,5-trien-8-oneas a yellow solid (16 mg, yield; 40%).

Molecular weight: 295.34; Mass spectrometry: 296 (M+H)⁺; In vitroactivity grade: B; Cellular activity grade: (A549)-B; ¹H-NMR (300 MHz,CDCl3-d): 1.42 (1H, d, J=14.3 Hz), 1.84 (1H, m), 1.92 (1H, d, J=13.6Hz), 2.17 (1H, m), 2.85 (1H, s), 3.16 (1H, td, J=3.0, 12.7 Hz), 3.35(1H, dd, J=1.9, 12.2 Hz), 3.63 (1H, d, J=13.2 Hz), 3.92 (1H, dd, J=4.5,12.8 Hz), 6.88 (1H, td, J=1.1, 8.3 Hz), 6.99 (1H, dd, J=1.1, 8.3 Hz),7.04 (1H, s), 7.32 (1H, td, J=1.1, 8.3 Hz), 7.75 (1H, dd, J=1.5, 8.0Hz).

Example 8-2

To a solution of2-amino-4-{1-[(benzyloxy)carbonyl]-3-piperidinyl}-6-(2-hydroxyphenyl)nicotinicacid (750 mg, 1.676 mmol), which was obtained in the step (2) of Example8-1, methylamine hydrochloride (230 mg, 3.352 mmol), triethylamine (340mg, 3.35 mmol), and 1-hydroxybenzotriazole (360 mg, 2.68 mmol), wasadded 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (390mg, 2.0 mmol) at 0° C. under an argon atmosphere. The mixture wasstirred at room temperature overnight. The reaction mixture wasextracted with ethyl acetate, washed with water and brine, dried overMgSO₄, filtered, and concentrated under reduced pressure. The residuewas purified by column chromatography on silica gel (Hexane:ethylacetate, 2:1) to give benzyl3-{2-amino-6-(2-hydroxyphenyl)-3-(N-methylcarbamoyl)-4-pyridinyl}-1-piperidinecarboxylateas a pale yellow solid (337 mg, yield; 44%).

To a solution of benzyl3-{2-amino-6-(2-hydroxyphenyl)-3-(N-methylcarbamoyl)-4-pyridinyl}-1-piperidinecarboxylate(130 mg, 0.274 mmol) in MeOH (2 mL) and THF (2 mL) was added Pd—C (180mg). The mixture was stirred at room temperature under a hydrogenatmosphere overnight. The reaction mixture was filtered on Celite®,washed with THF, and then concentrated under reduced pressure. Theresidue was diluted with ethyl acetate, and added 4N HCl in 1,4-dioxane.The resulting yellow precipitates were washed with acetonitrile. Thecollected solid was dried under reduced pressure to give2-amino-6-(2-hydroxyphenyl)-N-methyl-4-(3-piperidinyl)nicotinamidehydrochloride(10 mg, yield; 81%).

Molecular weight: 362.86; Mass spectrometry: 327 (M+H)⁺; In vitroactivity grade: B; Cellular activity grade: (A549)-B; ¹H-NMR (500 MHz,DMSO-d6): 1.69-1.77 (2H, m), 1.82-1.86 (2H, m), 2.80 (3H, d, J=4.4 Hz),2.88 (1H, m), 3.03 (1H, m), 3.19-3.29 (3H, m), 6.89-6.93 (3H, m),7.29-7.32 (2H, m), 7.85 (1H, br), 8.50 (1H, br), 8.85 (1H, br), 9.16(1H, br).

Example 8-3

With the use of the starting compound 1G (instead of 1D), 2C and othermaterials,2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-N-methyl-4-(3-piperidinyl)nicotinamidehydrochloride was prepared in the similar manner as that of Example 8-1and 8-2.

Molecular weight: 432.95; Mass spectrometry: 397 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (300 MHz,DMSO-d6): 0.29-0.34 (2H, m), 0.49-0.55 (2H, m), 1.21 (1H, m), 1.74-1.89(SH, m), 2.84 (3H, d, J=4.5 Hz), 3.10-3.11 (2H, m), 3.25-3.28 (2H, m),3.78-3.90 (2H, m), 6.59 (1H, d, J=8.3 Hz), 6.65 (1H, d, J=8.3 Hz), 7.20(1H, s), 7.26 (1H, t, J=8.3 Hz), 7.56 (1H, br s), 8.76 (1H, br s), 9.08(1H, br s), 13.8 (1H, br s).

Example 9-1

(1) A mixture of the starting compound 1A (3.500 g, 15.47 mmol),starting compound 2B (3.299 g, 15.47 mmol), tert-butyl cyanoacetate(2.184 g, 15.47 mmol), ammonium acetate (3.577 g, 46.40 mmol) and1,2-dimethoxyethane (17 mL) was heated at reflux for 3.5 hrs. Aftercooled to room temperature, the mixture was concentrated under reducedpressure, and the residue was partitioned between ethyl acetate andwater. The separated organic phase was washed with brine, dried overNa₂SO₄, filtered, and concentrated under reduced pressure. The residuewas purified by flash chromatography on silica gel (hexane: ethylacetate, 4:1) to give tert-butyl2-amino-6-[2-(benzyloxy)phenyl]-4-[1-(tert-butoxycarbonyl)-3-piperidinyl]nicotinate.(1.892 g, yield; 22%).

(2) Then benzyl moiety was removed in a similar manner as described inthe step (2) of Example 1-1. The resulting solid was suspended inethanol, collected by filtration, washed with ethanol, and dried underreduced pressure to give tert-butyl2-amino-4-[1-(tert-butoxycarbonyl)-3-piperidinyl]-6-(2-hydroxyphenyl)nicotinate(0.322 g, yield; 38%).

(3) tert-Butyl2-amino-4-[1-(tert-butoxycarbonyl)-3-piperidinyl]-6-(2-hydroxyphenyl)nicotinate(0.050 g, 0.106 mmol)) was treated under acidic conditions in a similarmanner as described in the step (3) of Example 1-1. to give tert-butyl2-amino-6-(2-hydroxyphenyl)-4-(3-piperidinyl)nicotinate hydrochloride(0.034 g, yield; 79%).

Molecular weight: 405.92; Mass spectrometry: 370 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-B; ¹H-NMR (500 MHz,DMSO-d6): 1.60 (9H, s), 1.76-1.95 (4H, m), 2.90-2.97 (1H, m), 3.20-3.46(4H, m), 6.89-6.94 (2H, m), 7.31-7.36 (1H, m), 7.38 (1H, s), 8.02 (1H,dd, J=1.3, 8.2 Hz), 9.09 (1H, br), 9.29 (1H, br), 13.78 (1H, br).

Example 9-2

To a solution of tert-butyl2-amino-6-(2-hydroxyphenyl)-4-(3-piperidinyl)nicotinate hydrochloride(0.023 g, 0.057 mmol), which was obtained in Example 9-1, in methylenechloride (1.0 mL) was added trifluoroacetic acid (TFA) (1.0 mL). Afterbeing stirred for 6 hrs, the mixture was concentrated under reducedpressure. The residue was dissolved in 1,4-dioxane (3.0 mL), and thentreated with a HCl solution of 1,4-dioxane (4N, 0.2 mL). The resultingprecipitates were collected by filtration under an argon atmosphere,washed with 1,4-dioxane and acetonitrile, and dried under reducedpressure to give 2-amino-6-(2-hydroxyphenyl)-4-(3-piperidinyl)nicotinicacid hydrochloride (0.020 g, yield; quant.).

Molecular weight: 349.82; Mass spectrometry: 314 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-B; ¹H-NMR (500 MHz,DMSO-d6): 1.75-1.93 (4H, m), 2.92 (1H, br m), 2.92-3.39 (3H, m),3.63-3.67 (1H, br m), 6.91-6.96 (2H, m), 7.32-7.36 (1H, m), 7.37 (1H,s), 7.95 (1H, d, J=7.9 Hz), 8.84 (1H, br), 9.41 (1H, br), 13.80 (1H,br).

Example 9-3

With the use of 1-(2-benzyloxy-6-cyclopropylmethoxy-phenyl)-ethanoneprepared in the similar manner as that of the starting compound 1G, thestarting compound 2B and other materials,2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-(3-piperidinyl)nicotinicacid hydrochloride was prepared in a similar manner as described inExample 9-2.

Molecular weight: 419.91; Mass spectrometry: 384 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-C; ¹H-NMR (500 MHz,DMSO-d6): 0.31-0.32 (2H, m), 0.51-0.53 (2H, m), 1.20 (1H, m), 1.70-1.74(2H, m), 1.89-1.91 (2H, m), 2.89 (1H, m), 3.18 (1H, m), 3.26-3.28 (3H,m), 3.83 (2H, dd, J=6.9, 7.3 Hz), 6.58 (1H, d, J=8.2 Hz), 6.65 (1H, d,J=8.5 Hz), 7.28 (1H, dd, J=8.2, 8.5 Hz), 7.30 (1H, s), 7.81 (1H, br s),8.92 (1H, br s), 9.37 (1H, br s).

Example 10-1

(1) To a solution of2-amino-6-[2-(benzyloxy)phenyl]-4-[1-(tert-butoxycarbonyl)-3-piperidinyl]nicotinicacid (0.500 g, 0.993 mmol) which was derived from the product in thestep (1) of Example 9-1, methylamine hydrochloride (0.134 g, 1.896mmol), triethylamine (0.277 mL, 1.986 mmol), and 1-hydroxybenzotriazole(0.215 g, 1.589 mmol) in dichloromathane (10.0 mL), at 0° C. under anargon atmosphere was added 1-ethyl-3-(3-dimethylaminopropyl)carbodiimidehydrochloride (0.228 g, 1.191 mmol). The mixture was stirred at roomtemperature overnight. The reaction mixture was concentrated underreduced pressure, then extracted with ethyl acetate and water. Theseparated organic phase was washed with brine, dried over MgSO₄,filtered, then concentrated under reduced pressure. The resultingresidue was purified by column chromatography on Silica-gel(dichloromathane/methanol=19/1) to give tert-butyl3-{2-amino-6-[2-(benzyloxy)phenyl]-3-[(methylamino)carbonyl]-4-pyridinyl}-1-piperidinecarboxylateas a yellow oil (0.11 g, 21%).

(2) To a cooled (−20° C.) solution of tert-butyl3-{2-amino-6-[2-(benzyloxy)phenyl]-3-[(methylamino)carbonyl]-4-pyridinyl}-1-piperidinecarboxylate(0.120 g, 0.232 mmol) and triethylamine (0.213 g, 2.090 mmol) intetrahydrofuran (10 mL) was added triphosgene (0.069 g, 0.232 mmol)under an argon atmosphere. The stirring was continued at 0° C. to roomtemperature overnight. The reaction mixture was extracted with ethylacetate and water. The separated organic phase was washed with brine,dried over MgSO₄, filtered, and concentrated under reduced pressure. Theresulting residue was purified by preparative TLC (hexane/ethylacetate=2/1) to give tert-butyl3-{7-[2-(benzyloxy)phenyl]-3-methyl-2,4-dioxo-1,2,3,4-tetrahydropyrido[2,3-d]pyrimidin-5-yl}-1-piperidinecarboxylateas a pale yellow solid (0.0655 g, 52%).

(3) tert-butyl3-{7-[2-(benzyloxy)phenyl]-3-methyl-2,4-dioxo-1,2,3,4-tetrahydropyrido[2,3-d]pyrimidin-5-yl}-1-piperidinecarboxylate(0.065 g, 0.120 mmol) was treated in a similar manner as that of thestep (2) of Example 1-1. The residue was purified by preparative TLC(hexane/ethyl acetate=1/1) to give tert-butyl3-[7-(2-hydroxyphenyl)-3-methyl-2,4-dioxo-1,2,3,4-tetrahydropyrido[2,3-d]pyrimidin-5-yl]-1-piperidinecarboxylate(0.010 g, 18%), which is then treated as described in the step (3) ofExample 1-1 to obtain7-(2-hydroxy-phenyl)-3-methyl-5-piperidin-3-yl-1H-pyrido[2,3-d]pyrimidine-2,4-dionehydrochloride.

Molecular weight: 388.86; Mass spectrometry: 353 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: A549-B; ¹H-NMR (500 MHz,DMSO-d6): 1.81-1.85 (2H, m), 1.96-1.98 (2H, m), 2.91 (1H, m), 3.26 (3H,s), 3.31-3.40 (3H, m), 4.70 (1H, m), 6.97-7.00 (2H, m), 7.39 (1H, m),7.86 (1H, s), 8.16 (1H, m), 8.73 (1H, br), 9.17 (1H, br), 12.37 (1H, s).

Example 10-2

With the use of 1-(2-benzyloxy-6-cyclopropylmethoxy-phenyl)-ethanoneprepared in the similar manner as that of the starting compound 1G, thestarting compound 2B and other materials,7-(2-Cyclopropylmethoxy-6-hydroxy-phenyl)-3-methyl-5-piperidin-3-yl-1H-pyrido[2,3-d]pyrimidine-2,4-dionewas prepared in a similar manner as that of Example 10-1.

Molecular weight: 458.95; In vitro activity grade: A; ¹H-NMR (500 MHz,DMSO-d6): 0.31-0.34 (2H, m), 0.56-0.60 (2H, m), 1.27 (1H, m), 1.67 (1H,m), 1.79 (1H, m), 1.91-2.01 (2H, m), 2.87 (1H, m), 3.06 (1H, m), 3.27(3H, s), 3.30 (1H, m), 3.39 (1H, m), 3.83 (1H, dd, J=7.3, 9.8 Hz), 3.91(2H, dd, J=6.9, 9.8 Hz), 4.65 (1H, m), 6.58 (2H, d, J=8.5 Hz), 7.25 (1H,dd, J=8.2, 8.5 Hz), 7.91 (1H, s), 8.83 (1H, br s), 9.32 (1H, br s), 12.2(1H, br s).

Example 11-1

(1) To a cold (0° C.) solution of tert-butyl2-amino-4-[1-(tert-butoxycarbonyl)-3-piperidinyl]-6-(2-hydroxyphenyl)nicotinate,obtained in the step (2) of Example 9-1, (0.200 g, 0.426 mmol) in THF(3.0 mL) under an argon atmosphere was added LiBH₄ (0.019 g, 0.85 mmol).After stirred at room temperature overnight, the mixture was quenchedwith water, and then extracted with ethyl acetate and 0.5N HCl. Theseparated organic phase was washed with water and brine, dried overNa₂SO₄, filtered and concentrated under reduced pressure. The residuewas purified by column chromatography on silica gel (hexane: ethylacetate, 1:1) to give tert-butyl3-[2-amino-3-(hydroxymethyl)-6-(2-hydroxyphenyl)-4-pyridinyl]-1-piperidinecarboxylate.(0.161 g, yield; 95%)

tert-Butyl3-[2-amino-3-(hydroxymethyl)-6-(2-hydroxyphenyl)-4-pyridinyl]-1-piperidinecarboxylate(0.160 g, 0.401 mmol) in 1,4-dioxane (3.0 mL) was treated under acidicconditions in a similar manner as described in the step (3) of Example1-1 to give2-[6-amino-5-(hydroxymethyl)-4-(3-piperidinyl)-2-pyridinyl]phenolhydrochloride. (0.132 g, yield; 98%).

Molecular weight: 335.84; Mass spectrometry: 300 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 1.75-1.98 (4H, m), 2.90 (1H, br), 3.14-3.33 (3H, m), 3.56-3.60(1H, br), 4.58 (2H, dd, J=13.0, 20.2 Hz), 6.96 (H, t, J=7.6 Hz), 7.06(1H, d, J=7.9 Hz), 7.19 (1H, s), 7.37 (1H, br t, J=7.3 Hz), 7.65 (1H, brd, J=7.6 Hz), 9.03 (1H, br), 9.32 (1H, br), 13.59 (1H, br).

Example 11-2

With the use of starting compound 1I and 2B, and other materials,2-[6-amino-5-(hydroxymethyl)-4-(3-piperidinyl)-2-pyridinyl]-3-fluorophenolhydrochloride was prepared in a similar manner as that of Example 11-1.

Molecular weight: 353.83; Mass spectrometry: 318 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 1.03 (3H, t, J=7.6 Hz), 1.83 (4H, m), 3.00 (2H, m), 3.31 (2H,m), 3.71 (1H, m), 4.56 (2H, dd, J=13.1, 19.4), 5.16 (1H, br), 6.79 (2H,m), 7.12 (1H, s), 7.32 (1H, m), 8.52 (1H, d, J=9.8 Hz), 8.86 (1H, d,J=9.8 Hz), 14.0 (1H, br).

Example 11-3

With the use of the starting compound 1F, the starting compound 2B, andother materials,6′-amino-5′-(hydroxymethyl)-4′-(3-piperidinyl)-2,2′-bipyridin-3-olhydrochloride was prepared in a similar manner as that of Example 11-1.

Molecular weight: 336.82; Mass spectrometry: 301 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 1.81-1.95 (3H, m), 2.46-2.64 (1H, m), 3.00-3.06 (2H, m),3.28-3.39 (2H, m), 3.54-3.60 (1H, m), 4.56 (1H, d, J=13.0 Hz), 4.59 (1H,d, J=13.0 Hz), 7.41-7.50 (2H, m), 7.81 (1H, s), 8.24 (1H, d, J=3.8 Hz),8.92 (1H, br), 9.07 (1H, br).

Example 11-4

(1) With the use of the starting compound 1D instead of 1A and 2Hinstead of 2B, di(tert-butyl)2′-amino-6′-{2-[(4-methoxybenzyl)oxy]phenyl}-5,6-dihydro-3,4′-bipyridine-1,3′(2H)-dicarboxylatewas prepared in a similar method as that of step (1) of Example 9-1.

(2) To a cooled (0° C.) solution of di(tert-butyl)2′-amino-6′-{2-[(4-methoxybenzyl)oxy]phenyl}-5,6-dihydro-3,4′-bipyridine-1,3′(2H)-dicarboxylate(50.0 mg, 0.09 mmol) in THF (4.5 mL) was added sodiumbis(2-methoxyethoxy)aluminum hydride (0.10 mL) dropwise, then stirredfor 1 hr at 0° C. The reaction was quenched by an addition of saturatedNH₄Cl solution, filtered, and washed with ethyl acetate. The combinedorganic phase was washed with brine, dried over MgSO₄, filtered andevaporated. The crude product was purified by preparative silica gel TLC(15% acetone in chloroform) to give tert-butyl2′-amino-3′-(hydroxymethyl)-6′-{2-[(4-methoxybenzyl)oxy]phenyl}-5,6-dihydro-3,4′-bipyridine-1(2H)-carboxylateas a white foam (25 mg, yield 57%).

(3) tert-butyl2′-amino-3′-(hydroxymethyl)-6′-{2-[(4-methoxybenzyl)oxy]phenyl}-5,6-dihydro-3,4′-bipyridine-1(2H)-carboxylate(25.0 mg, 0.05 mmol) was treated with 2N HCl in dioxane (4.0 mL)overnight at room temperature. The resulting solid was collected byfiltration, washed with ether and dried under reduced pressure to give2-[6′-amino-5′-(hydroxymethyl)-1,2,5,6-tetrahydro-3,4′-bipyridin-2′-yl]phenolhydrochloride. (14 mg, yield 87%).

Molecular weight: 333.82; Mass spectrometry: 298 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (300 MHz,D2O): 2.51 (2H, br s), 3.35 (2H, br s), 3.83 (2H, s), 4.58 (2H, s), 6.04(1H, br s), 6.93-7.01 (3H, m), 7.34-7.47 (2H, m).

Example 12-1

(1) A mixture of 1-[2,6-bis(benzyloxy)phenyl]ethanone (8.00 g, 24.067mmol) (starting compound 1E), tert-butyl3-formyl-1-piperidinecarboxylate (5.133 g, 24.067 mmol) (startingcompound 2B), tert-butylcyanoacetate (3.398 g, 24.067 mmol), andammonium acetate (5.432 g, 72.202 mmol) in 1,2-dimethoxyethane (24 mL)was stirred at 100° C. in a sealed tube overnight. After cooled to roomtemperature, the reaction mixture was partitioned between ethyl acetateand water. The separated organic phase was washed with brine, dried overMgSO₄, filtered and concentrated under reduced pressure. The residue wasdiluted with 30 mL of dichloromethane, and MnO₂ was added to themixture, and then the mixture was stirred at room temperature for 5 hrs.The mixture was filtered on Celite®, and concentrated under reducedpressure. The resulting residue (liquid) was purified by columnchromatography on silica gel (hexane/ethyl acetate=2/1) to givetert-butyl2-amino-6-[2,6-bis(benzyloxy)phenyl]-4-[1-(tert-butoxycarbonyl)-3-piperidinyl]nicotinateas a pale yellow oil. (2.5 g, yield; 16%)

(2) A suspension of tert-butyl2-amino-6-[2,6-bis(benzyloxy)phenyl]-4-[1-(tert-butoxycarbonyl)-3-piperidinyl]nicotinate(1.00 g, 1.052 mmol) and 10% Paradium-carbon (0.500 g) in 20 mL of ethylacetate was stirred at room temperature under a hydrogen atmosphere (1atm) overnight. After paradium carbon was removed by filtration onCelite®, the filtrate was concentrated under reduced pressure to givetert-butyl2-amino-4-[1-(tert-butoxycarbonyl)-3-piperidinyl]-6-(2,6-dihydroxyphenyl)nicotinateas a brown oil. (0.710 g, yield; 97%)

(3) To a solution of tert-butyl2-amino-4-[1-(tert-butoxycarbonyl)-3-piperidinyl]-6-(2,6-dihydroxyphenyl)nicotinate(1.280 g, 2.636 mmol) and K₂CO₃ (3.643 g, 26.36 mmol) in DMF (100 mL)was added (bromomethyl)cycropropane (0.268 mL, 2.768 mmol). The stirringwas continued at 50° C. for 20 hrs. After the solvent was removed byevaporation, the residue was extracted with ethyl acetate and water. Theseparated organic phase was washed with brine, dried over MgSO₄,filtered, then concentrated under reduced pressure. The resultingresidue was purified by column chromatography on Silica-gel(hexane/ethylacetate=3/1) to give tert-butyl2-amino-4-[1-(tert-butoxycarbonyl)-3-piperidinyl]-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]nicotinateas a brown solid (0.820 g, yield; 58%).

tert-butyl2-amino-4-[1-(tert-butoxycarbonyl)-3-piperidinyl]-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]nicotinatewas treated in a similar manner as that of the step (1) of Example 11-1and then in a similar manner as that of the step (3) of Example 1-1 toobtain2-[6-amino-5-(hydroxymethyl)-4-(3-piperidinyl)-2-pyridinyl]-3-(cyclopropylmethoxy)phenolhydrochloride.

Molecular weight: 405.93; Mass spectrometry: 370 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (300 MHz,DMSO-d6): 0.26-0.29 (2H, m), 0.47-0.52 (2H, m), 1.13 (1H, m), 1.75-1.83(5H, m), 2.90-2.93 (2H, m), 3.04-3.17 (2H, m), 3.81-3.86 (2H, m), 4.60(2H, s), 6.55 (1H, d, J=8.3 Hz), 6.60 (1H, d, J=8.3 Hz), 7.06 (1H, s),7.27 (1H, t, J=8.3 Hz), 7.59 (1H, br), 8.96 (1H, br), 9.25 (1H, br),13.58 (1H, br).

Examples 12-2 to 12-7

According to the similar synthetic procedure of Example 12-1, compoundsshown in Table 4 were prepared.

TABLE 4 Mol In Ex. No Structure weight Mass vitro A549 NMR 12-02

441.96 406 A B (500 MHz, DMSO-d6): 1.56-1.88 (4H, m), 2.84-3.20(3H, m),4.54 (1H, d, J=13.2Hz), 4.57(1H, d, J=13.2Hz), 5.08(2H, s), 6.70 (1H, d,J=8.2Hz), 6.76(1H, d, J= 8.5Hz),7.05(1H, s), 7.29- 7.41(6H, m), 7.60(1H,br), 8.95 (1H, br), 9.25(1H, br), 10.42(1H, br), 13.67(1H, br). 12-03

393.92 358 A A (500 MHz, DMSO-d6): 0.87(3H, t, J=7.2Hz), 1.64(2H, q,J=6.9 Hz), 1.88-1.90(3H, m), 2.07 (1H, m), 2.88(1H, m), 3.05(1H, m),3.22-3.24(2H, m), 3.56(1H, m), 3.92(2H, t, J=6.6Hz), 4.59 (2H, d,J=3.1Hz), 4.96(1H, br), 6.62(1H, d, J=8.2Hz), 6.66(1H, d, J=8.2Hz),7.00(1H, s), 7.29 (1H, t, J=8.2Hz), 7.58(1H, # br), 8.92(1H, br),9.21(1H, br), 13.57 (1H, br). 12-04

421.97 386 A A (500 MHz, DMSO-d6): 0.83(9H, s), 0.82-1.05(4H,m), 1.75-1.93(4H, m), 2.87(1H, br), 3.02- 3.07(4H, m), 4.61(2H, dd, J= 13.2,18.6Hz), 4.99(1H, br), 6.60 (1H, d, J=8.5Hz), 6.67(1H, d, J= 8.2Hz),6.79-6.89(1H, m), 6.98(1H, s), 7.07(1H, br), 7.27- 7.29(1H, br),7.62(1H, br), 13.72 (1H, br). 12-05

405.93 370 A A (500 MHz, DMSO-d6): 0.29-0.32 (2H, m), 0.46-0.50(2H, m),1.19 (1H, m), 1.89-2.05(2H, m), 3.01- 3.07(2H, m), 3.34-3.48(3H, m),3.84(2H, d, J=6.9Hz), 4.61 (2H, s), 6.59(1H, d, J=8.5Hz), 6.69(1H, d,J=8.2Hz), 6.80(1H, s), 7.27(1H, dd, J=8.2, 8.5Hz), 7.75(1H, br s),9.14-9.25(2H, m), 13.6(1H, br). 12-06

393.92 358 A A (500 MHz, DMSO-d6): 0.85(3H, t, J=7.3Hz), 1.63-1.67(2H,m), 1.82-1.85(2H, m), 1.90-1.95 (2H, m), 3.03-3.08(2H, m), 3.34-3.45(4H, m), 3.92(2H, t, J=6.6 Hz), 4.60(2H, s), 6.62(1H, d, J= 8.2Hz),6.68(1H, d, J=8.2Hz), 6.75(1H, s), 7.28(1H, t, J=8.2 Hz), 7.63(1H, br),8.98(2H, br). 12-07

436.00 400 A A (500 MHz, DMSO-d6): 0.79-0.82 (7H, m), 1.12-1.16(2H, m),1.42- 1.49(1H, m), 1.60-1.66(2H, m), 1.82-1.84(2H, m), 1.93- 1.98(2H,m), 3.93-3.95(2H, m), 4.60(2H, s), 6.61(1H, d, J=8.2 Hz), 6.68(1H, d,J=8.2Hz), 6.74 (1H, s), 7.28(1H, t, J=8.2Hz), 7.68(2H, s), 9.07(2H, s),10.38 (1H, brs).

Example 13-1

(1) To a cold (0° C.) solution of tert-butyl2-amino-6-[2-(benzyloxy)phenyl]-4-[1-(tert-butoxycarbonyl)-3-piperidinyl]nicotinate,which was obtained in the step (1) of Example 9-1, (2.590 g, 4.627 mmol)in THF (25 mL) was added LiBH₄ (0.202 g, 9.255 mmol). The mixture wasstirred at room temperature for 5 hrs, and then quenched with water. Themixture was extracted with ethyl acetate and water. The separatedorganic phase was washed with water and brine, dried over Na₂SO₄,filtered and concentrated under reduced pressure. The residue waspurified by column chromatography on silica gel (hexane: ethyl acetate,2:3) to give tert-butyl3-[2-amino-6-[2-(benzyloxy)phenyl]-3-(hydroxymethyl)-4-pyridinyl]-1-piperidine-carboxylate.(0.904 g, yield; 40%)

(2) To a solution of tert-butyl3-[2-amino-6-[2-(benzyloxy)phenyl]-3-(hydroxymethyl)-4-pyridinyl]-1-piperidinecarboxylate(0.900 g, 1.838 mmol) in methylene chloride (20 mL) was added manganese(IV) oxide (3.20 g, 36.8 mmol). After stirred at room temperature for 40min, the mixture was filtered to remove the manganese salt. The filtratewas concentrated under reduced pressure. The resulting solid waspurified by recrystallization from a mixture of ethyl acetate and hexaneto give tert-butyl3-[2-amino-6-[2-(benzyloxy)phenyl]-3-formyl-4-pyridinyl]-1-piperidinecarboxylate.(0.651 g, yield; 73%)

(3) Then the benzyl moiety was removed in a similar manner as describedin the step (2) of Example 1-1. The residue was purified by columnchromatography on silica gel (hexane: ethyl acetate, 2:1) to givetert-butyl3-[2-amino-3-formyl-6-(2-hydroxyphenyl)-4-pyridinyl]-1-piperidinecarboxylate.(0.536 g, yield; quant.)

(4) To a solution of tert-butyl3-[2-amino-3-formyl-6-(2-hydroxyphenyl)-4-pyridinyl]-1-piperidinecarboxylate(0.100 g, 0.252 mmol) in 1,4-dioxane (2.0 mL) under an argon atmospherewas added a HCl solution of 1,4-dioxane (4N, 2.0 mL), and the stirringwas continued at room temperature for 1 hr. The mixture was diluted withEt₂O, and the supernatant was decanted. The precipitates were washedtwice with Et₂O, and dried under reduced pressure to give a pale yellowsolid, which was then dissolved in methanol (2.0 mL). The mixture wasallowed to cool with ice-water bath. Then sodium cyanoborohydride (0.047g, 0.76 mmol) was added to the mixture under an argon atmosphere. Themixture was allowed to warm to room temperature, and the stirring wascontinued overnight. The resulting mixture was concentrated underreduced pressure. The residue was dissolved in a mixture of methylenechloride (2.0 mL) and THF (2.0 mL). To the mixture was addedtriethylamine (0.140 mL, 1.006 mmol) followed by di-tert-butyldicarbonate (0.110 g, 0.503 mmol), and the stirring was continued for 2hrs. The resulting mixture was partitioned between ethyl acetate andwater. The separated organic phase was washed with brine, dried overNa₂SO₄, filtered, and concentrated under reduced pressure. The residuewas purified by column chromatography on silica gel (hexane:ethylacetate, 2:1) to give tert-Butyl3-[2-amino-6-(2-hydroxyphenyl)-3-(methoxymethyl)-4-pyridinyl]-1-piperidinecarboxylate.(0.033 g, yield; 47%)

(4) tert-Butyl3-[2-amino-6-(2-hydroxyphenyl)-3-(methoxymethyl)-4-pyridinyl]-1-piperidinecarboxylate(0.025 g, 0.060 mmol) was treated under acidic conditions in a similarmanner as described in the step (3) of Example 1-1 to give2-[6-amino-5-(methoxymethyl)-4-(3-piperidinyl)-2-pyridinyl]phenolhydrochloride.

Molecular weight: 349.86; Mass spectrometry: 314 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 1.40-1.74 (4H, m), 2.80-2.92 (2H, m), 4.03-4.13 (2H, m), 4.42(1H, d, J=11.5 Hz), 4.50 (1H, d, J=11.5 Hz), 6.22-6.32 (2H, m),6.81-6.85 (2H, m), 7.17 (1H, s), 7.21 (1H, br t, J=7.3 Hz), 7.93 (1H, brd, J=7.6 Hz), 14.30 (1H, br).

Example 14-1

To a cold (0° C.) solution of tert-butyl3-[2-amino-6-[2-(benzyloxy)phenyl]-3-(hydroxymethyl)-4-pyridinyl]-1-piperidinecarboxylate(0.104 g, 0.212 mmol) obtained in the step (1) of Example 13-1 in THF(2.0 mL) including triethylamine (0.089 mL, 0.64 mmol) was addedtriphosgene (0.044 g, 0.15 mmol). After stirred for 30 min, the mixturewas allowed to warm to room temperature, and the stirring was continuedfor further 1 hr. The resulting mixture was quenched with water andextracted with ethyl acetate. The separated organic phase was washedwith brine, dried over Na₂SO₄, filtered and concentrated under reducedpressure. The residue was purified by column chromatography on silicagel (hexane:ethyl acetate, 5:3) to give tert-butyl3-{7-[2-benzyloxy]phenyl]-2-oxo-1,4-dihydro-2H-pyrido[2,3-d][1,3]-oxazin-5-yl}-1-piperazinecarboxylate.(0.058 g, yield; 53%)

A solution of tert-butyl3-{7-[2-benzyloxy]phenyl]-2-oxo-1,4-dihydro-2H-pyrido-[2,3-d][1,3]oxazin-5-yl}-1-piperazinecarboxylate(0.050 g, 0.097 mmol) in ethyl acetate (3.0 mL) was hydrogenated at 1atm in the presence of palladium on charcoal (10%, 0.10 g) overnight.The resulting mixture was filtered on Celite® and washed with ethylacetate and THF. The combined filtrate was concentrated under reducedpressure, and the residue was purified by column chromatography onsilica gel (hexane:ethyl acetate, 2:1) to give tert-Butyl3-[2-amino-6-(2-hydroxyphenyl)-3-methyl-4-pyridinyl]-1-piperidinecarboxylate.(0.027 g, yield; 73%)

tert-Butyl3-[2-amino-6-(2-hydroxyphenyl)-3-methyl-4-pyridinyl]-1-piperidinecarboxylate(0.025 g, 0.065 mmol) was treated under acidic conditions in a similarmanner as described in the step (3) of Example 1-1 to give2-[6-amino-5-methyl-4-(3-piperidinyl)-2-pyridinyl]phenol hydrochloride.(0.017 g, yield; 82%).

Molecular weight: 319.84; Mass spectrometry: 284 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A/Jurkat-B; ¹H-NMR‘(500 MHz, DMSO-d6): 1.08-1.75 (4H, m), 2.10 (3H, s), 2.55-2.98 (3H, m),4.12 (1H, br), 6.19 (2H, br d, J=15.1 Hz), 6.79-6.83 (2H, m), 7.12 (1H,s), 7.18 (1H, br t, J=7.9 Hz), 7.88 (1H, br).

Example 15-1

(1) To a cold (−20° C.) solution of tert-butyl3-{2-amino-6-[2-(benzyloxy)phenyl]-3-formyl-4-pyridinyl}-1-piperidinecarboxylate(400 mg, 0.82 mmol), which was obtained in the step (2) of Example 13-1,in THF (2.0 mL) was added dropwise a solution methyl magnesium bromidein THF (7.38 mL, 7.38 mol). The mixture was stirred at −20° C. for 2hrs, quenched with saturated aqueous NH₄Cl solution, and extracted withethyl acetate. The separated organic phase was washed with water andbrine, dried over Na₂SO₄, filtered and concentrated under reducedpressure. The residue was purified by silica gel column chromatography(hexane/ethyl acetate, 3:2) to give tert-butyl3-[2-amino-6-[2-(benzyloxy)phenyl]-3-(1-hydroxyethyl)-4-pyridinyl]-1-piperidinecarboxylateas a yellow oil. (356 mg, yield; 86%)

(2) Then benzyl moiety was removed in a similar manner as described inthe step (2) of Example 1-1 to give tert-Butyl3-[2-amino-3-(1-hydroxyethyl)-6-(2-hydroxyphenyl)-4-pyridinyl]-1-piperidinecarboxylateas a pale yellow oil. (147 mg, yield; 96%)

(3) tert-Butyl3-[2-amino-3-(1-hydroxyethyl)-6-(2-hydroxyphenyl)-4-pyridinyl]-1-piperidinecarboxylate(30 mg, 0.07 mmol) was treated under acidic conditions in a similarmanner as described in the step (3) of Example 1-1 to give2-[6-amino-5-(1-hydroxyethyl)-4-(3-piperidinyl)-2-pyridinyl]phenolhydrochloride as a yellow solid. (20 mg, yield; 78%).

Molecular weight: 349.86; Mass spectrometry: 296 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-B; ¹H-NMR (500 MHz,DMSO-d6): 1.83-1.92 (7H, m), 2.90-2.91 (1H, m), 3.13-3.15 (1H, m), 3.57(3H, s), 5.29-5.39 (1H, m), 6.89-6.99 (1H, m), 7.05-7.10 (1H, m), 7.17(1H, d, J=3.16 Hz), 7.27-7.40 (2H, m), 7.62 (1H, d, J=5.0 Hz), 7.68-7.87(2H, m), 13.50 (1H, br s).

Example 15-2

In a similar manner as that of Example 15-1,2-[6-amino-5-[hydroxy(phenyl)methyl]-4-(3-piperidinyl)-2-pyridinyl]phenolhydrochloride was prepared.

Molecular weight: 411.94; Mass spectrometry: 376 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-B; ¹H-NMR (500 MHz,DMSO-d6): 1.67-1.74 (2H, m), 1.89-1.99 (2H, m), 2.88-2.93 (1H, m),3.14-3.24 (4H, m), 6.32 (1H, s), 6.91 (1H, d, J=2.2 Hz), 6.92-7.01 (2H,m), 7.24-7.32 (3H, m), 7.34-7.40 (6H, m), 7.42-7.44 (1H, m), 7.75 (1H,d, J=7.9 Hz).

Example 15-3

To a stirred solution of tert-butyl3-[2-amino-6-[2-(benzyloxy)phenyl]-3-(1-hydroxyethyl)-4-pyridinyl]-1-piperidinecarboxylate(0.100 g, 0.199 mmol) obtained in the step (1) of Example 15-1 indichrolomethane (1 mL) was added MnO₂ (0.350 g, 3.971 mmol). The mixturewas stirred at room temperature for 2 hrs. The mixture was filtered onCelite® and concentrated under reduced pressure to give tert-butyl3-{3-acetyl-2-amino-6-[2-(benzyloxy)phenyl]-4-pyridinyl}-1-piperidinecarboxylate.

tert-butyl3-{3-acetyl-2-amino-6-[2-(benzyloxy)phenyl]-4-pyridinyl}-1-piperidinecarboxylate (0.090 g, 0.169 mmol) was then treated in a similar manneras that of the step (2) of Example 1-1. The residue was recrystallizedfrom ethyl alcohol to give tert-butyl3-[3-acetyl-2-amino-6-(2-hydroxyphenyl)-4-pyridinyl]-1-piperidinecarboxylateas a pale yellow solid. (0.019 g, yield 27%)

Then tert-butyl3-[3-acetyl-2-amino-6-(2-hydroxyphenyl)-4-pyridinyl]-1-piperidinecarboxylate(0.020 g, 0.044 mmol) was treated in a similar manner as that of thestep (3) of Example 1-1. The resulting precipitate was collected byfiltration, washed with acetonitril, and dried under reduced pressure togive1-[2-amino-6-(2-hydroxyphenyl)-4-(3-piperidinyl)-3-pyridinyl]ethanonehydrochloride. (0.014 g, yield 89%).

Molecular weight: 347.85; Mass spectrometry: 312 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO): 1.17-1.87 (4H, m), 2.92-2.98 (1H, m), 2.56 (3H, s), 3.00-3.29(4H, m), 6.98-6.91 (4H, m), 7.30 (1H, dd, J=7.3, 7.9 Hz), 7.37 (1H, s),8.73 (1H, d, J=7.6 Hz), 13.73 (1H, br s).

Example 16-1

To a solution of copper (II) bromide (0.82 g, 3.692 mmol) inacetonitrile (10 mL) was added tert-butyl nitrate (0.550 mL, 4.614mmol). The mixture was stirred at 65° C. for 15 min. A solution oftert-butyl3-{2-amino-6-[2-(benzyloxy)phenyl]-3-formyl-4-pyridinyl}-1-piperidinecarboxylate(1.50 g, 3.076 mmol), which was obtained in the step (2) of Example13-1, in acetonitrile (10 mL) was added to the mixture, and stirred at65° C. for 3 hrs. After cooled to room temperature, the reaction mixturewas diluted with water and extracted with ethyl acetate. The organicphase was dried over sodium sulfate, filtered and concentrated underreduced pressure. The resulting residue was purified by silica gelcolumn chromatography (hexane/ethyl acetate=4/1-2/1-1/1) to givetert-butyl3-{6-[2-(benzyloxy)phenyl]-2-bromo-3-formyl-4-pyridinyl}-1-piperidinecarboxylateas a white solid (0.260 g, yield; 15%).

To a stirred solution of tert-butyl3-{6-[2-(benzyloxy)phenyl]-2-bromo-3-formyl-4-pyridinyl}-1-piperidinecarboxylate(0.150 g, 0.272 mmol) in 1,4-dioxane (1 mL) was added hydrazinemonohydrate (0.5 mL). The reaction mixture was stirred at 90° C. for 15hrs. After cooled to room temperature, the reaction was quenched withwater and extracted with ethyl acetate. The organic phase was dried oversodium sulfate, filtered and concentrated under reduced pressure. Theresidue was recrystallized from diethyl ether to give tert-butyl3-{6-[2-(benzyloxy)phenyl]-1H-pyrazolo[3,4-b]pyridin-4-yl}-1-piperidinecarboxylateas a white solid (0.030 g, 23%). Then tert-butyl3-{6-[2-(benzyloxy)phenyl]-1H-pyrazolo[3,4-b]pyridin-4-yl}-1-piperidinecarboxylatewas treated in a similar manner as that of the step (2) of Example 1-1and then the step (3) of Example 1-1 to obtain2-(4-Piperidin-3-yl-1H-pyrazolo[3,4-b]pyridin-6-yl)-phenolhydrochloride.

Molecular weight: 330.82; Mass spectrometry: 295 (M+H)⁺; In vitroactivity grade: B; Cellular activity grade: (A549)-B; ¹H-NMR (500 MHz,DMSO-d6): 1.91-2.05 (4H, m), 6.97-7.00 (2H, m), 7.35-7.87 (1H, m), 7.87(1H, s), 8.16 (1H, d, J=7.3 Hz), 8.42 (1H, s), 8.96 (1H, br s), 9.26(1H, br s).

Example 17-1

(1) A mixture of the starting compound 1G (10.00 g, 30.638 mmol), 2B(13.069 g, 61.275 mmol), tert-butylcyanoacetate (8.650 g, 61.275 mmol),and ammonium acetate (6.902 g, 91.913 mmol) in dioxane (10 mL) wasstirred at 90° C. overnight. After cooled to room temperature, thereaction mixture was diluted with ethyl acetate (100 mL). To the mixturewas added chloranil (1.507 g, 6.128 mmol), and stirred at roomtemperature. After 1.5 hrs, ascorbic acid (1.079 g, 6.128 mmol) wasadded to the mixture. After stirred for 1.5 hrs, the mixture waspartitioned between ethyl acetate and water. The organic phase waswashed with brine, dried over MgSO₄, filtered, and then concentratedunder reduced pressure. The resulting residue was purified by columnchromatography on Silica-gel (hexane/ethyl acetate=2/1) to givetert-butyl2-amino-4-[1-(tert-butoxycarbonyl)-3-piperidinyl]-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}nicotinateas a pale brown form (4.9 g, 24%)

(2) To a cooled solution of tert-butyl2-amino-4-[1-(tert-butoxycarbonyl)-3-piperidinyl]-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}nicotinate(4.9 g, 7.426 mmol) in tetrahydrofuran (60 mL) was added dropwiseVitride® (10 mL) under an argon atmosphere. The stirring was continuedat 0° C. for 1 hr. After quenched by saturated aqueous NH₄Cl solution,saturated aqueous potassium sodium tartrate was added to the mixture,then the mixture was stirred vigorously. The mixture was extracted withethyl acetate, washed with water and brine, dried over MgSO₄, filtered,and concentrated under reduced pressure to give tert-butyl3-[2-amino-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-3-(hydroxymethyl)-4-pyridinyl]-1-piperidinecarboxylate,which was used for the next step without further purification (4.38 g,yield; quant.).

(3) To a stirred solution of tert-butyl3-[2-amino-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-3-(hydroxymethyl)-4-pyridinyl]-1-piperidinecarboxylate(0.501 g, 0.850 mmol) in 1,4-dioxane was added 37% formaldehyde solution(5.000 mL) and 1N HCl (5.000 mL). The mixture was stirred at roomtemperature for 2 hrs. The reaction mixture was extracted with ethylacetate and water. The separated organic phase was washed with brine,dried over Na₂SO₄, filtered and concentrated under reduced pressure. Theresulting residue was purified by column chromatography on silica gel(ethyl acetate/hexane=3/2) to give 1,1-dimethylethyl(tert-butyl)-3-{7-[2-[(cyclopropylmethyl)oxy]-6-({[4-(methyloxy)phenyl]methyl}oxy)phenyl]-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-5-yl}-1-piperidinecarboxylateas a colorless foam (0.319 g, yield; 62%).

(4) To a stirred solution of1,1-dimethylethyl(tert-butyl)-3-{7-[2-[(cyclopropylmethyl)oxy]-6-({[4-(methyloxy)phenyl]methyl}oxy)phenyl]-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-5-yl}-1-piperidinecarboxylate(0.050 g, 0.083 mmol) in 1,4-Dioxane was added 4N HCl in 1,4-dioxane(2.000 mL). The mixture was stirred at room temperature for 6 hrs. Thereaction mixture was concentrated under reduced pressure. The resultingresidue was washed with acetonitrile to give3-(cyclopropylmethoxy)-2-[5-(3-piperidinyl)-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-7-yl]-phenolhydrochloride as a colorless solid (0.033 g, yield; 95%).

Molecular weight: 417.94; Mass spectrometry: 382 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 0.30 (2H, br s), 0.51 (2H, br s), 1.17 (1H, m), 1.84 (4H, m),2.86 (1H, m), 4.91 (2H, br), 5.01 (2H, br), 6.58 (1H, d, J=7.9 Hz), 6.64(1H, br), 7.16 (1H, br), 7.26 (1H, t, J=7.9 Hz), 8.30 (1H, br), 9.14(1H, br), 9.29 (1H, br), 13.95 (1H, br).

Example 18-1

To a cooled (0° C.) solution of tert-butyl3-[2-amino-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-3-(hydroxymethyl)-4-pyridinyl]-1-piperidinecarboxylate(5.0 g, 8.478 mmol), which was obtained in the step (2) of Example 17-1,and diisopropylethyl amine (4.12 mL, 25.435 mmol) in tetrahydrofuran(200 mL) under argon atmosphere was added dropwise to a solution oftriphosgene (1.258 g, 4.239 mmol) in tetrahydrofuran (100 mL). Themixture was allowed to warm to room temperature, and the stirring wascontinued for 3 hrs. After quenched by water, the mixture was extractedwith ethyl acetate. The separated organic phase was washed with brine,dried over MgSO₄, filtered, and concentrated under reduced pressure. Theresulting residue was purified by column chromatography on Silica-gel(hexane/ethyl acetate=1/1) to give tert-butyl3-(7-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-2-oxo-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-5-yl)-1-piperidinecarboxylateas a white form (3.2 g, yield; 61%).

To a solution of tert-butyl3-(7-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-2-oxo-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-5-yl)-1-piperidinecarboxylate(2.0 g, 3.248 mmol) in dioxane (15 mL) was added 4N HCl in dioxane (30mL) at room temperature. The stirring was continued for 3 hrs. After thesolvent was removed by evaporation, the resulting solid was trituratedwith acetonitril, collected by filtration, and washed with acetonitrile.The solid was dried under reduced pressure to give7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-5-(3-piperidinyl)-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-2-onehydrochloride as a white solid (0.865 g, yield; 62%).

Molecular weight: 431.92; Mass spectrometry: 396 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 0.26-0.37 (2H, m), 0.51-0.63 (2H, m), 1.20-1.31 (1H, m),1.72-1.95 (4H, m), 2.80-2.96 (2H, m), 3.17-3.37 (3H, m), 3.79-3.88 (2H,m), 5.48 (1H, d, J=14.2 Hz), 5.53 (1H, d, J=14.2 Hz), 6.54 (2H, d, J=8.2Hz), 7.17 (1H, t, J=8.2 Hz), 7.77 (1H, s), 9.05 (1H, br), 9.27 (1H, br),10.96 (1H, s).

Example 18-2

To a cold (0° C.) solution of tert-butyl3-[2-amino-3-(hydroxymethyl)-6-(2-hydroxyphenyl)-4-pyridinyl]-1-piperidinecarboxylate(0.297 g, 0.74 mmol), which was obtained in the step (1) of Example 11-1in THF (3.00 mL) including triethylamine (0.226 g, 2.23 mmol) was addedtriphosgene (0.221 g, 0.74 mmol). After 30 min, the mixture was allowedto warm to room temperature and the stirring was continued for 1 hr. Thereaction was quenched with an aqueous NaHCO₃ solution, and extractedwith ethyl acetate. The organic phase was washed with aqueous NH₄Clsolution, water, and brine successively. The organic phase was driedover Na₂SO₄, filtered and concentrated under reduced pressure. Theresulting residue was purified by silica gel column chromatography(hexane/ethyl acetate, 3:2) to give tert-butyl3-[7-(2-hydroxyphenyl)-2-oxo-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-5-yl]-1-piperidinecarboxylateas a pale yellow solid. (46.2 mg, yield; 14.6%)

To a solution of tert-butyl3-[7-(2-hydroxyphenyl)-2-oxo-1,4-dihydro-2H-pyrido-[2,3-d][1,3]oxazin-5-yl]-1-piperidinecarboxylate(45 mg, 0.11 mmol) in 1,4-dioxane (1.0 mL) was added 4N HCl in1,4-dioxane (2.0 mL). The mixture was stirred for 1 hr at roomtemperature. The resulting solid was collected by filtration, washedwith 1,4-dioxane, and dried under reduced pressure to give7-(2-hydroxyphenyl)-5-(3-piperidinyl)-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-2-onehydrochloride as a white solid. (22.8 mg, yield; 60%).

Molecular weight: 361.83; Mass spectrometry: 326 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A /Jurkat-B; ¹H-NMR(500 MHz, DNSO-d6): 1.80-1.96 (5H, m), 2.86-2.93 (1H, m), 3.13-3.26 (3H,m), 5.46-5.55 (2H, m), 6.91-6.94 (2H, m), 7.30-7.33 (1H, m), 7.77 (1H,s), 8.03-8.04 (1H, m), 11.15 (1H, s), 12.46 (1H, br s).

Example 18-3

With the use of tert-butyl3-[2-amino-3-(1-hydroxyethyl)-6-(2-hydroxyphenyl)-4-pyridinyl]-1-piperidinecarboxylate,which was obtained in the step (2) of Example 15-1,7-(2-Hydroxy-phenyl)-4-methyl-5-piperidin-3-yl-1,4-dihydro-benzo[d][1,3]-oxazin-2-onehydrochloride was prepared in a similar manner as that of Example 18-2.

Molecular weight: 375.86; Mass spectrometry: 340 (M+H)⁺; In vitroactivity grade: B; Cellular activity grade: (A549)-B; ¹H-NMR (500 MHz,DMSO-d6): 1.52-1.53 (3H, m), 1.80-2.04 (5H, m), 2.91-2.95 (1H, m),3.13-3.21 (3H, m), 5.83-5.85 (1H, m), 6.92-6.95 (3H, m), 7.32 (1H, dd,J=7.9, 7.3 Hz), 8.04 (1H, d, J=7.6 Hz), 11.2 (1H, s), 12.44 (1H, br s).

Example 18-4/18-5

A mixture of the starting compound 1A (3.000 g, 13.258 mmol),N-tert-butoxycarbonyl-phenylalaninal (3.305 g, 13.258 mmol), tert-butylcyanoacetate (1.872 g, 13.258 mmol), ammonium acetate (3.066 g, 39.774mmol) and 1,2-dimethoxyethane (5.0 mL) was heated at reflux for 6 hrs.After cooled to room temperature, the mixture was extracted with ethylacetate and saturated NaHCO₃ solution. The separated organic phase waswashed with water and brine successively, dried over Na₂SO₄, filteredand concentrated under reduced pressure. The residue was triturated withdiisopropyl ether to give tert-butyl2-amino-6-[2-(benzyloxy)phenyl]-4-{1-[(tert-butoxycarbonyl)amino]-2-phenylethyl}-1,4-dihydro-3-pyridinecarboxylateas a yellow solid (1.281 g, yield; 16%).

To a solution of tert-butyl2-amino-6-[2-(benzyloxy)phenyl]-4-{1-[(tert-butoxycarbonyl)amino]-2-phenylethyl}-1,4-dihydro-3-pyridinecarboxylate(1.200 g, 2.007 mmol) in methylene chloride (20.0 mL) at roomtemperature was added chloranil (0.543 g, 2.208 mmol), and the stirringwas continued for 1 hr. The mixture was filtered and the filtrate wasconcentrated under reduced pressure. The residue was extracted withethyl acetate and 10% aqueous NaHCO₃ solution. The separated organicphase was washed with water and brine successively, dried over Na₂SO₄,filtered and concentrated under reduced pressure. The crude product wascrystallized from ethanol to give tert-butyl2-amino-6-[2-(benzyloxy)phenyl]-4-{1-[(tert-butoxycarbonyl)amino]-2-phenylethyl}nicotinateas a pale yellow solid (0.972 g, yield; 84%).

To a cold (0° C.) solution of tert-butyl2-amino-6-[2-(benzyloxy)phenyl]-4-{1-[(tert-butoxycarbonyl)amino]-2-phenylethyl}nicotinate(0.100 g, 0.198 mmol) in THF (1.50 mL) was added dropwise Vitride®(0.500 mL), and the stirring was continued for 2 hrs. The mixture wasquenched at 0° C. with saturated aqueous NH₄Cl solution, and extractedwith ethyl acetate. The separated organic phase was washed with waterand brine successively, dried over Na₂SO₄, filtered and concentratedunder reduced pressure. The crude product was purified by columnchromatography on silica gel (hexane:ethyl acetate, 1:1) to givetert-butyl-1-{2-amino-6-[2-(benzyloxy)phenyl]-3-formyl-4-pyridinyl}-2-phenylethylcarbamateas a colorless oil (0.044 g, yield; 51%).

To a solution of tert-butyl1-{2-amino-6-[2-(benzyloxy)phenyl]-3-formyl-4-pyridinyl}-2-phenylethylcarbamate(0.040 g, 0.092 mmol) in ethanol (1.0 mL) was added NaBH₄ (0.010 g, 0.26mmol), and the stirring was continued for 3 hrs. The mixture wasextracted with ethyl acetate and saturated aqueous ammonium chloridesolution. The separated organic phase was washed with water and brinesuccessively, dried over Na₂SO₄, filtered and concentrated under reducedpressure. The crude product was crystallized from ethanol to givetert-butyl1-[2-amino-6-[2-(benzyloxy)phenyl]-3-(hydroxymethyl)-4-pyridinyl]-2-phenylethylcarbamateas a pale yellow solid (0.023 g, yield; 57%).

To a cold (0° C.) solution of tert-butyl1-[2-amino-6-[2-(benzyloxy)phenyl]-3-(hydroxymethyl)-4-pyridinyl]-2-phenylethylcarbamate(0.210 g, 0.400 mmol) in THF (3.0 mL) including triethylamine (0.167 mL,1.199 mmol) was added dropwise a solution of triphosgene (0.059 g, 0.200mmol) in THF (1.0 mL). After stirred at 0° C. for 45 min, the mixturewas allowed to warm to room temperature, and the stirring was continuedfor 30 min. The reaction mixture was cooled at 0° C., and then quenchedwith saturated aqueous NaHCO₃ solution. The resulting mixture wasextracted with ethyl acetate and water. The separated organic phase waswashed with brine, dried over Na₂SO₄, filtered and concentrated underreduced pressure. The crude product was purified by columnchromatography on silica gel (hexane:ethyl acetate, 2:1) to givetert-butyl1-{7-[2-(benzyloxy)phenyl]-2-oxo-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-5-yl}-2-phenylethylcarbamateas a colorless oil (0.170 g, yield; 77%).

Then in a similar manner as that of the step (2) of Example 1-1 and thestep (3) of Example 1-1, tert-butyl1-{7-[2-(benzyloxy)phenyl]-2-oxo-1,4-dihydro-2H-pyrido-[2,3-d][1,3]oxazin-5-yl}-2-phenylethylcarbamatewas treated to give5-(1-Amino-2-phenyl-ethyl)-7-(2-hydroxy-phenyl)-1,4-dihydro-pyrido[2,3-d][1,3]oxazin-2-onehydrochloride.

Example 18-4/18-5

Molecular weight: 397.86; Mass spectrometry: 362 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-B; ¹H-NMR (500 MHz,DMSO-d6): 3.11 (1H, dd, J=10.1, 12.9 Hz), 3.44 (1H, dd, J=5.0, 12.9 Hz),4.25 (1H, d, J=14.5 Hz), 4.71-4.74 (1H, m), 5.35 (1H, d, J=14.5 Hz),6.93-6.99 (2H, m), 7.08 (2H, dd, J=1.9, 7.3 Hz), 7.23-7.28 (3H, m),7.33-7.36 (1H, m), 8.13 (1H, dd, J=1.3, 7.9 Hz), 8.37 (1H, s), 8.98 (3H,br), 11.18 (1H, s), 12.32 (1H, br).

Example 18-6

To a cooled (0° C.) solution of tert-butyl2′-amino-3′-(hydroxymethyl)-6′-{2-[(4-methoxybenzyl)oxy]phenyl}-5,6-dihydro-3,4′-bipyridine-1(2H)-carboxylate(25.0 mg, 0.05 mmol), which was obtained in the step (2) of Example11-4, and triethylamine (0.02 mL, 0.14 mmol) in THF (3.0 mL) was added asolution of triphosgene (5.7 mg, 0.02 mmol) in THF (3.0 mL), and thestirring was continued for 30 min. The reaction mixture was allowed towarm to room temperature. The reaction was cooled to 0° C. and thenquenched by an addition of saturated aqueous NaHCO₃ solution, extractedwith ethyl acetate. The separated organic phase was washed with brine,dried over MgSO₄, filtered and evaporated. The crude product waspurified by preparative silica gel TLC (10% acetone in chloroform) togive tert-butyl5-(7-{2-[(4-methoxybenzyl)oxy]phenyl}-2-oxo-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-5-yl)-3,6-dihydro-1(2H)-pyridinecarboxylateas a white solid. (21.7 mg, yield 83%).

Then the tert-butyl and methoxy benzyl were removed in a similar manneras that of the steo(2) of Example 1-1.

Molecular weight: 359.82; Mass spectrometry: 324 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,MeOH-d4): 2.63-2.65 (2H, m), 3.44-3.49 (2H, m), 4.02 (2H, br d, J=1.9Hz), 5.45 (2H, s), 6.04 (1H, s), 6.88-6.95 (2H, m), 7.29-7.32 (1H, m),7.63 (1H, s), 7.90 (1H, dd, J=1.3, 7.9 Hz).

Examples 18-7 to 18-16

According to the similar synthetic procedure of Examples 18-1 to 18-6,compounds shown in Table 5 were prepared.

TABLE 5 Mol In Ex. No Structure weight Mass vitro A549 NMR 18-07

379.82 no peak A A (500 MHz, DMSO-d6): 1.82(4H, m), 3.02(3H, m),3.29(2H, d, J= 21.5Hz), 5.50(2H, dd, J=23.9, 39.6Hz), 6.73(1H, d, J=23.9Hz), 6.78(1H, m), 7.28(1H, dd, J= 13.6, 24.8Hz), 7.34(1H, s), 8.58(1H,m), 8.97(1H, m), 10.95(1H, s). 18-08

362.82 327 A A (500 MHz, DMSO-d6): 1.88-1.94 (4H, m), 3.26-3.33(2H, m),3.57- 3.59(1H, m), 3.63-3.74(2H, m), 5.51(1H, d, J=14.2Hz), 5.58(1H, d,J=14.2Hz), 7.39- 7.40(2H, m), 8.14(1H, s), 8.22 (1H, t, J=2.8Hz),11.25(1H, s). 18-09

326.36 327 C A (500 MHz, CDCl3): 0.34-0.44 (2H, m), 0.61-0.72(2H, m),1.24- 1.35(1H, m), 2.33(3H, s), 3.89 (2H, d, J=6.9Hz), 5.39(2H, s),6.45(1H, dd, J=1.0, 8.5Hz), 6.65(1H, dd, J=1.0, 8.5Hz), 7.18(1H, t,J=8.5Hz), 7.54(1H, br s), 8.10(1H, s), 12.48(1H, br s). 18-10

467.96 432 A A (500 MHz, DMSO-d6): 1.25-1.32 (1H, m), 1.53-1.83(3H, m),2.58- 2.80(2H, m), 3.01-3.07(1H, m), 3.13-3.18(1H, m), 3.27- 3.30(1H,m), 5.07(2H, dd, J= 11.0, 18.2Hz), 5.44(2H, dd, J= 14.2, 24.0Hz),6.58(1H, d, J= 8.2Hz), 6.72(1H, d, J=8.2Hz), 7.22(1H, t, J=8.2Hz), 7.34-7.46(6H, m), 8.77(1H, br), # 9.04 (1H, br), 10.92(1H, s). 18-11

419.91 384 A A (300 MHz, DMSO-d6): 0.90(3H, t, J=7.2Hz), 1.72(2H, m),1.84- 1.88(2H, m), 2.85-2.92(2H, m), 3.18-3.34(4H, m), 3.90-3.95 (2H,m), 5.48(2H, d, J=5.6Hz), 6.55(2H, dd, J=8.3, 8.5Hz), 7.17(1H, t,J=8.3Hz), 7.54(1H, s), 8.88(1H, br), 9.13(1H, br), 10.89(1H, s). 18-12

377.83 342 B C (500 MHz, DMSO-d6): 1.70-1.73 (2H, m), 1.78-1.89(2H, m),2.87- 2.90(2H, m), 3.00-3.05(2H, m), 4.05(1H, m), 5.49(2H, s), 6.39(1H,d, J=8.2Hz), 6.42 (1H, d, J=8.2Hz), 7.04(1H, m), 7.92(1H, m), 8.55(1H,br), 8.77 (1H, br), 10.95(1H, s), 10.99(1H, s). 18-13

431.92 396 A A (500 MHz, DMSO-d6): 0.28-0.30 (2H, m), 0.55-0.58(2H, m),1.31 (1H, m), 1.88(4H, m), 3.01-3.04 (3H, m), 3.35-3.38(2H, m), 3.84(2H, d, J=6.9Hz), 5.50(2H, s), 6.53(2H, d, J=8.2Hz), 7.17(1H, t,J=8.2Hz), 7.74(1H, s), 8.92 (1H, br), 9.00(1H, br), 10.95(1H, s). 18-14

419.91 384 A A (500 MHz, DMSO-d6): 0.90(3H, t, J=7.5Hz), 1.75(2H, m),1.85 (4H, m), 3.02(3H, m), 3.36(2H, m) 3.95(2H, t, J=7.6Hz), 5.49 (2H,s), 6.53(1H, d, J=7.9Hz), 6.57(1H, d, J=8.2Hz), 7.18(1H, t, J=8.2Hz),7.50(1H, s), 8.88 (1H, br), 8.99(1H, br), 10.90(1H, s). 18-15

461.99 426 A A (500 MHz, DMSO-d6): 0.81(6H, d, J=6.6Hz), 1.14-1.19(2H,m), 1.47-1.51(1H, m), 1.67- 1.71(2H, m), 1.83-1.90(4H, m), 2.96-3.06(3H,m), 3.31-3.33 (2H, m), 3.95-3.97(2H, m), 5.49 (2H, s), 6.53-6.58(2H, m),7.17 (1H, t, J=8.3Hz), 7.42(1H, s), 9.05-9.11(1H, m), 10.88(1H, s).18-16

545.52 A B (500 MHz, DMSO-d6): 1.56-1.64 (4H, m), 2.95(4H, br s),5.09(2H, s), 5.45(2H, s), 6.55-6.59(1H, m), 6.70(1H, d, J=8.3Hz), 7.20(1H, t, J=8.1Hz), 7.33-7.38 (6H, m), 8.26(1H, br s), 8.57 (1H, br s),10.85(1H, br s), 11.11 (1H, br s).

Example 19-1

A mixture of7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-5-(3-piperidinyl)-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-2-onehydrochloride (0.070 g, 0.160 mmol), which was obtained in Example 18-1,and 2.5N HCl in 1,4-dioxane (4 mL) was stirred at 80° C. overnight.After cooled to room temperature, the mixture was concentrated underreduced pressure. The resulting residue was diluted with dichloromethane(3 mL), and triethylamine (0.064 g, 0.497 mmol) was added. To the cooled(0° C.) mixture was added di-tert-butyl dicarbonate (0.047 g, 0.218mmol) under an argon atmosphere. The stirring was continued at roomtemperature overnight. After quenched by water, the mixture wasextracted with ethyl acetate and water. The separated organic phase waswashed with brine, dried over MgSO₄, filtered, and concentrated underreduced pressure. The residue was purified by preparative TLC(hexane/ethyl acetate=1/1) to give tert-butyl3-[7-(2,6-dihydroxyphenyl)-2-oxo-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-5-yl]-1-piperidine-carboxylate(0.019 g, 19%). To a solution of tert-butyl3-[7-(2,6-dihydroxyphenyl)-2-oxo-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-5-yl]-1-piperidinecarboxylatein dioxane (1 mL) was added 4N HCl in dioxane (1 mL) at roomtemperature. The stirring was continued for 3 hrs. After the solvent wasremoved by evaporation, the resulting residue was washed withacetonitril, and dried under reduced pressure to give7-(2,6-dihydroxyphenyl)-5-(3-piperidinyl)-1,4-dihydro-2H-pyrido[2,3-d][1,3]-oxazin-2-onehydrochloride as a brown solid (0.007 g, 46%).

Molecular weight: 377.83; Mass spectrometry: 342 (M+H)⁺; In vitroactivity grade: Cellular activity grade: (A549)-; ¹H-NMR (500 MHz,DMSO-d6): 1.73-1.89 (4H, m), 2.89-2.94 (2H, m), 3.19 (1H, m), 3.25-3.32(2H, m), 5.46 (1H, d, J=14.2 Hz), 5.51 (1H, d, J=14.2 Hz), 6.41 (2H, d,J=8.2 Hz), 7.03 (1H, t, J=8.2 Hz), 7.88 (1H, s), 8.90 (1H, br), 9.14(1H, br), 11.01 (1H, s).

Example 20-1

To a stirred solution of tert-butyl3-[2-amino-6-[2-(benzyloxy)-6-hydroxyphenyl]-3-(hydroxymethyl)-4-pyridinyl]-1-piperidinecarboxylate(0.100 g, 0.198 mmol), which was obtained in the similar procedure asthat of Example 13-1, in dichrolomethane (3 mL) was added MnO₂ (0.340 g,3.956 mmol). The mixture was stirred at room temperature for 2 hrs. Themixture was filtered on Celite® and concentrated under reduced pressure.The residue was recrystallized from ethyl alcohol to give tert-butyl3-{2-amino-6-[2-(benzyloxy)-6-hydroxyphenyl]-3-formyl4-pyridinyl}-1-piperidinecarboxylateas a yellow solid. (0.092 g, yield 93%)

To a stirred solution of tert-butyl3-{2-amino-6-[2-(benzyloxy)-6-hydroxyphenyl]-3-formyl-4-pyridinyl}-1-piperidinecarboxylate(0.090 g, 0.183 mmol) in 1,4-dioxane (2 mL) was added 4N HCl in dioxane(2 mL). The mixture was stirred at room temperature for 2 hrs. Theresulting precipitate was collected by filtration, washed withacetonitril, and dried under reduced pressure to give2-amino-6-[2-(benzyloxy)-6-hydroxyphenyl]-4-(3-piperidinyl)nicotinaldehydehydrochloride. (0.103 g, yield quant.)

To a cold (0° C.) solution of2-amino-6-[2-(benzyloxy)-6-hydroxyphenyl]-4-(3-piperidinyl)nicotinaldehydehydrochloride (0.100 g, 0.227 mmol) in methanol was added NaBH₃CN (0.040mL, 0.682 mmol) under an argon atmosphere. The mixture was stirred atroom temperature for 12 hrs and concentrated under reduced pressure. Theresidue was extracted with ethyl acetate and water. The separatedorganic phase was washed with saturated aqueous NaHCO₃ solution andbrine, dried over Na₂SO₄, filtered and concentrated under reducedpressure. The resulting residue was purified by column chromatography onsilica gel (hexane/ethyl acetate=2/1) to give2-(6-amino-5,9-diazatricyclo[7.3.1.02,7]trideca-2,4,6-trien-4-yl)-3-(benzyloxy)phenolas a white solid.

To a stirred solution of2-(6-amino-5,9-diazatricyclo[7.3.1.02,7]trideca-2,4,6-trien-4-yl)-3-(benzyloxy)phenolin 1,4-dioxane (2 mL) was added 4N HCl in dioxane (2 mL). The mixturewas stirred at room temperature for 2 hrs. The resulted precipitate wascollected by filtration, washed with acetonitril, and dried underreduced pressure to give2-(6-amino-5,9-diazatricyclo[7.3.1.02,7]trideca-2,4,6-trien-4-yl)-3-(benzyloxy)phenolhydrochloride.(0.018 g, yield 19%).

Molecular weight: 423.95; Mass spectrometry: 388 (M+H)⁺; In vitroactivity grade: B; Cellular activity grade: (A549)-C; ¹H-NMR (500 MHz,DMSO-d6): 1.10-1.23 (2H, m), 1.41 (1H, d, J=12.3 Hz), 1.71-1.80 (1H, m),2.79 (1H, d, J=12.9 Hz), 2.89-2.91 (2H, m), 2.97 (1H, d, J=12.9 Hz),3.44 (2H, d, J=18.3 Hz), 3.81 (1H, d, J=18.0 Hz), 5.09-5.17 (2H, m),6.01 (1H, s), 6.64 (1H, d, J=8.5 Hz), 7.13 (1H, t, J=8.2 Hz), 7.33-7.41(4H, m), 10 7.49 (2H, d, J=1.6 Hz).

Example 20-2

A mixture of5-[(1R)-1-amino-2-phenylethyl]-7-(2-hydroxyphenyl)-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-2-onehydrochloride (0.014 g, 0.035 mmol), which was obtained in Example 18-4,in acetonitrile (3.0 mL) was heated at reflux for 2 hrs. The resultingmixture was concentrated under reduced pressure. The residue wasdissolved in 1,4-dioxane (1.0 mL), treated with 4N HCl solution in1,4-dioxane (0.5 mL), and concentrated under reduced pressure. Theresidue was triturated with acetonitrile, and dried under reducedpressure to give2-[(1R)-4-amino-1-benzyl-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-6-yl]phenolhydrochloride as a white solid (0.010 g, yield; 80%).

Molecular weight: 353.85; Mass spectrometry: 318 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-B; ¹H-NMR (500 MHz,DMSO-d6): 3.30 (1H, dd, J=9.1, 14.1 Hz), 3.54 (1H, dd, J=5.0, 14.1 Hz),4.26-4.42 (2H, br m), 5.20 (1H, br), 6.91 (1H, t, J=7.9 Hz), 6.97 (1H,d, J=8.2 Hz), 7.29-7.39 (2H, m), 7.41 (1H, t, J=7.9 Hz), 7.62 (1H, d,J=7.3 Hz), 10.14 (1H, br), 10.31 (1H, br), 13.90 (1H, br).

Example 20-3

In a similar manner as described in Examples 20-1 and20-2,2-[(1S)-4-amino-1-benzyl-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-6-yl]phenolhydrochloride was prepared.

Molecular weight: 353.85; Mass spectrometry: 318 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-B; ¹H-NMR (500 MHz,DMSO-d6): 3.30 (1H, dd, J=9.1, 14.1 Hz), 3.54 (1H, dd, J=5.0, 14.1 Hz),4.26-4.42 (2H, br m), 5.20 (1H, br), 6.91 (1H, t, J=7.9 Hz), 6.97 (1H,d, J=8.2 Hz), 7.29-7.39 (2H, m), 7.41 (1H, t, J=7.9 Hz), 7.62 (H, d,J=7.3 Hz), 10.14 (1H, br), 10.31 (1H, br), 13.90 (1H, br).

Example 21-1

(1) To a stirred solution of tert-butyl3-[2-amino-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-3-(hydroxymethyl)-4-pyridinyl]-1-piperidinecarboxylate(0.050 g, 0.085 mmol), which was obtained in the step (2) of Example17-1, and phenyl disulfide (0.055 g, 0.254 mmol) in tetrahydrofuran (3mL) was added tributyl phosphine (0.060 mL, 0.254 mmol). The mixture wasstirred at room temperature for 24 hrs. The mixture was extracted withethyl acetate and water. The separated organic phase was washed withbrine, dried over Na₂SO₄, filtered, and concentrated under reducedpressure. The resulting residue (liquid) was purified by columnchromatography on silica gel (hexane/ethyl acetate=3/1-1/1) to givetert-butyl3-{2-amino-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-3-[(phenylsulfanyl)methyl]4-pyridinyl}-1-piperidinecarboxylate(0.021 g, yield; 37%) as a form.

(2) To a stirred solution of tert-butyl3-{2-amino-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-3-[(phenylsulfanyl)methyl]-4-pyridinyl}-1-piperidinecarboxylatein 1,4-dioxane (2 mL) was added 4N HCl in dioxane (2 mL). The mixturewas stirred at room temperature overnight, and diluted with ethylacetate. The resulting precipitate was collected by filtration, washedwith ethanol, and dried under reduced pressure to give2-[6-amino-5-[(phenylsulfanyl)methyl]4-(3-piperidinyl)-2-pyridinyl]-3-(cyclopropylmethoxy)phenolhydrochloride.

Molecular weight: 498.09; Mass spectrometry: 462 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-B; ¹H-NMR (500 MHz,DMSO-d6): 0.30 (2H, br s), 0.51 (2H, br s), 1.67 (2H, m), 1.80 (1H, br),4.26 (1H, d, J=12.6 Hz), 4.37 (1H, br), 6.59 (2H, br), 7.36 (2H, t,J=7.6 Hz), 7.46 (2H, d, J=7.6 Hz), 7.72 (1H, br), 8.61-8.98 (2H, br),10.36 (1H, br), 13.58 (1H, br).

Example 21-2

(1) With the use of the starting compound 1G, tert-butyl4-formyl-piperidine-1-carboxylic acid prepared in a similar manner asthat of the starting compound 2B, and other materials, tert-butyl4-[2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxy-phenyl]-3-(hydroxymethyl)-4-pyridinyl]-1-piperidinecarboxylatewas prepared in a similar manner as that of the step (1) of Example11-1.

(2) To a stirred solution of tert-butyl4-[2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-3-(hydroxymethyl)-4-pyridinyl]-1-piperidinecarboxylate(0.250 g, 0.528 mmol) in acetonitrile(10 mL) was added1,1′-carbonyldiimidazole (0.170 g, 1.056 mmol) at room temperature.After stirred for 2 hrs, the reaction mixture was extracted with ethylacetate and water. The separated organic phase was washed with brine,dried over Na₂SO₄, filtered, and concentrated under reduced pressure.The resulting solid was collected by filtration, washed with hexane, anddried under reduced pressure to give tert-butyl4-[2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-3-(1H-imidazol-1-ylmethyl)-4-pyridinyl]-1-piperidinecarboxylate.(0.242 g, yield 88%)

(3) To a stirred solution of tert-butyl4-[2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-3-(1H-imidazol-1-ylmethyl)-4-pyridinyl]-1-piperidinecarboxylate(0.260 g, 0.500 mmol) in 1,4-dioxane (10 mL) was added 4N HCl in dioxane(10 mL). The mixture was stirred at room temperature for 2 hrs. Theresulting precipitate was collected by filtration, washed withacetonitrile, and dried under reduced pressure to give2-[6-amino-5-(1H-imidazol-1-ylmethyl)-4-(4-piperidinyl)-2-pyridinyl]-3-(cyclopropylmethoxy)phenolhydrochloride. (0.248 g, yield; quant

Molecular weight: 455.99; Mass spectrometry: 420 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 0.32 (2H, d, J=4.7 Hz), 0.52 (2H, d, J=4.7 Hz), 1.27 (1H, brs), 1.71 (2H, d, J=13.6 Hz), 1.91-1.98 (2H, m), 3.00-3.31 (2H, m),3.32-3.35 (3H, m), 3.86 (2H, d, J=6.9 Hz), 5.62(2H, s), 6.58 (1H, d,J=8.5 Hz), 6.64 (1H, d, J=6.9 Hz), 7.26 (1H, dd, J=8.5, 7.9 Hz), 7.66(1H, s), 7.71 (1H, s), 9.31 (1H, s).

Example 21-3

With the use of diphenyl carbonate instead of 1,1′-carbonyldiimidazole,2-[6-amino-5-(phenoxymethyl)-4-(4-piperidinyl)-2-pyridinyl]-3-(cyclopropylmethoxy)phenolhydrochloride was prepared in a similar manner as that of Example 21-2.

Molecular weight: 482.03; Mass spectrometry: 446 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 0.31 (2H, d, J=4.7 Hz), 0.50 (2H, d, J=6.9 Hz), 1.16-1.19 (1H,m), 1.84 (2H, d, J=13.2 Hz), 1.91-1.99 (2H, m), 2.96 (2H, m), 3.57 (1H,s), 3.76 (2H, d, J=6.6 Hz), 5.20 (2H, s), 6.61 (1H, d, J=8.2 Hz), 6.66(1H, d, J=6.0 Hz), 6.75 (1, d, J=8.5 Hz), 7.20 (1H, t, J=7.3 Hz), 7.08(2H, d, J=7.9 Hz), 7.15 (1H, t, J=7.9 Hz), 7.36 (2H, t, J=7.9 Hz).

Example 214-4

With the use of N,N′-disuccinimidyl carbonate instead of1,1′-carbonyldiimidazole,1-{[2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-(4-piperidinyl)-3-pyridinyl]methoxy}-2,5-pyrrolidinedionehydrochloride was prepared in a similar manner as that of Example 21-2.

Molecular weight: 503.00; Mass spectrometry: 467 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 0.19 (2H, d, J=5.0 Hz), 0.52 (2H, d, J=7.3 Hz), 1.21 (1H, brs), 1.91-1.96 (4H, m), 2.71 (4H, s), 3.03 (2H, d, J=8.2 Hz), 3.85 (2H,d, J=6.9 Hz), 5.18 (2H, s), 6.59 (1H, d, J=8.5 Hz), 6.64 (1H, d, J=7.6Hz), 7.05 (1H, br s), 7.27 (1H, t, J=7.9 Hz), 7.60 (1H, br s), 8.82 (1H,br s), 8.92 (1H, d, J=9.1 Hz).

Example 22-1

(1) To a cold (0° C.) solution of tert-butyl3-[2-amino-6-[2-(benzyloxy)phenyl]-3-(hydroxymethyl)-4-pyridinyl]-1-piperidinecarboxylate(0.050 g, 0.102 mmol), obtained in the step (1) of Example 13-1, intoluene (1.0 mL) was added diphenylphosphoryl azide (0.024 mL, 0.112mmol) followed by 1,8-diazabicyclo-[5.4.0.]undec-7-ene (0.017 mL, 0.112mmol). After 30 min, the mixture was allowed to warm to roomtemperature, and the stirring was continued for 2 hrs. The mixture wasconcentrated under reduced pressure, and the residue was partitionedbetween ethyl acetate and water. The separated organic phase was washedwith brine, dried over Na₂SO₄, filtered and concentrated under reducedpressure. The residue was purified by column chromatography on silicagel (hexane: ethyl acetate, 2:1) to give tert-butyl3-{2-amino-3-(azidomethyl)-6-[2-(benzyloxy)phenyl]-4-pyridinyl}-1-piperidinecarboxylate.(0.038 g, yield; 72%)

(2) A solution of tert-butyl3-{2-amino-3-(azidomethyl)-6-[2-(benzyloxy)phenyl]-4-pyridinyl}-1-piperidinecarboxylate(0.030 g, 0.058 mmol) in ethyl acetate (1.0 mL) was hydrogenated at 1atm in the presence of palladium on charcoal (10%, 0.015 g) overnight.The resulting mixture was filtered and washed with ethyl acetate. Thecombined filtrate was concentrated under reduced pressure to givetert-butyl3-[2-amino-3-(aminomethyl)-6-(2-hydroxyphenyl)-4-pyridinyl]-1-piperidinecarboxylate,which was used for the next step without further purification.

(3) To a cold (0° C.) solution of tert-butyl3-[2-amino-3-(aminomethyl)-6-(2-hydroxyphenyl)-4-pyridinyl]-1-piperidinecarboxylate(0.020 g, 0.050 mmol) in THF (2.0 mL) under an argon atmosphere wasadded 1,1′-carbonyldiimidazole (0.009 g, 0.055 mmol). After stirred for15 min, the mixture was quenched with saturated sodium bicarbonatesolution. The resulting mixture was partitioned between ethyl acetateand saturated ammonium chloride solution. The separated organic phasewas washed with water and brine, dried over Na₂SO₄, filtered andconcentrated under reduced pressure. The residue was dissolved in ethylacetate (2.0 mL). The mixture was heated at 60° C. for 1 hr and allowedto cool to room temperature. The resulting precipitates were collectedby filtration, washed with ethyl acetate and dried under reducedpressure to give tert-Butyl3-[7-(2-hydroxyphenyl)-2-oxo-1,2,3,4-tetrahydropyrido[2,3-d]pyrimidin-5-yl]-1-piperidinecarboxylate.(0.007 g, yield; 33%)

(4) tert-Butyl3-[7-(2-hydroxyphenyl)-2-oxo-1,2,3,4-tetrahydropyrido[2,3-d]-pyrimidin-5-yl]-1-piperidinecarboxylate(0.006 g, 0.014 mmol) was treated under acidic conditions in a similarmanner as described in the step (3) of Example 1-1 to give7-(2-hydroxyphenyl)-5-(3-piperidinyl)-3,4-dihydropyrido[2,3-d]pyrimidin-2(1H)-onehydrochloride. (0.005 g, yield; 98%)

Molecular weight: 360.85; Mass spectrometry: 325 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: B; ¹H-NMR (500 MHz,DMSO-d6): 1.76-1.93 (4H, m), 2.88-3.37 (5H, m), 4.47 (2H, dd, J=15.4,24.3 Hz), 6.87-6.92 (2H, m), 7.26-7.30 (2H, m), 7.63 (1H, s), 8.00 (1H,d, J=8.2 Hz), 8.57 (1H, br), 9.01 (1H, br), 10.08 (1H, s), 12.78 (1H,br).

Example 22-2

With the use of the starting compound 1G, 2B and other materials,7-[2-(cyclo-propylmethoxy)-6-hydroxyphenyl]-5-(3-piperidinyl)-3,4-dihydro-1H-pyrido[2,3-d]-pyrimidin-2-onehydrochloride was prepared in a similar manner as that of Example 22-1.

Molecular weight: 430.94; Mass spectrometry: 395 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 0.34 (2H, m), 0.57 (2H, m), 1.27 (1H, m), 1.73-1.87 (4H, m),2.82-2.90 (2H, m), 3.14 (1H, m), 3.29-3.33 (2H, m), 3.80-3.88 (2H, m),4.49 (2H, s), 6.52 (2H, d, J=8.2 Hz), 7.16 (1H, t, J=8.2 Hz), 7.75 (1H,s), 9.11 (1H, br), 9.26 (1H, br), 9.92 (1H, s).

Example 22-3

With the use of the starting compound 1G and tert-butyl4-formyl-piperidine-1-carboxylic acid and other materials,7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-5-(4-piperidinyl)-3,4-dihydro-1H-pyrido[2,3-d]pyrimidin-2-onehydrochloride was prepared in a similar manner as that of Example 22-1.

Molecular weight: 430.94; Mass spectrometry: 395 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 0.32 (2H, d, J=4.5 Hz), 0.57 (2H, d, J=1.7 Hz), 1.33-1.38 (1H,m), 1.80-1.91 (4H, m), 2.96-3.05 (3H, m), 3.34-3.39 (3H, m), 4.47 (2H,s), 6.51 (2H, dd, J=3.8, 8.3 Hz), 7.15 (1H, t, J=7.9 Hz), 7.24 (1H, s),7.74 (1H, s), 9.86 (1H, s).

Example 22-4

With the use of tert-butyl3-[2-amino-3-(aminomethyl)-6-(2-hydroxyphenyl)-4-pyridinyl]-1-piperidinecarboxylateobtained in the step (2) of Example 22-1,7-(2-hydroxyphenyl)-5-(3-piperidinyl)-3,4-dihydro-1H-pyrido[2,3-d]pyrimidine-2-thione hydrochloride was prepared in a similarmanner as that of Example 22-1 or 22-2.

Molecular weight: 447.00; Mass spectrometry: 411 (M+H)⁺; In vitroactivity grade: Cellular activity grade: (A549)-; ¹H-NMR (500 mHz,DMSO-d6): 0.33-0.36 (2H, m), 0.57-0.59 (2H, m), 1.28 (1H, m), 1.72-1.81(2H, m), 1.89-1.90 (2H, m), 2.82-2.89 (2H, m), 3.00 (1H, m), 3.27-3.34(2H, m), 3.82-3.85 (2H, m), 4.54-4.56 (2H, m), 6.52 (2H, d, J=8.2 Hz),7.16 (1H, t, J=8.2 Hz), 7.83 (1H, s), 8.73 (1 H, br), 9.08 (2H, br),11.13 (1H, s).

Example 22-5

A mixture of tert-butyl3-[2-amino-3-(aminomethyl)-6-(2-hydroxyphenyl)-4-pyridinyl]-1-piperidinecarboxylate(0.060 g, 0.151 mmol), which was obtained in the step (2) of Example22-1, sulfamine (0.022 g, 0.226 mmol) and pyridine (2.0 mL) was stirredat reflux overnight. After cooled to room temperature, the mixture waspartitioned between ethyl acetate and water. The separated organic phasewas washed with 0.5N HCl solution, water and brine successively, driedover Na₂SO₄, filtered and concentrated under reduced pressure. The crudeproduct was purified by column chromatography on silica gel (hexane:ethyl acetate, 1:1) to give tert-butyl3-[7-(2-hydroxyphenyl)-2,2-dioxido-3,4-dihydro-1H-pyrido[2,3-c][1,2,6]thiadiazin-5-yl]-1-piperidinecarboxylateas a pale yellow foam (0.059 g, yield; 85%).

Then the tert-butyl carbamate was treated in a similar manner as that ofthe step (3) of Example 1-1 to give2-[2,2-dioxido-5-(3-piperidinyl)-3,4-dihydro-1H-pyrido-[2,3-c][1,2,6]thiadiazin-7-yl]phenolhydrochloride.

Molecular weight: 396.90; Mass spectrometry: 361 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-B; ¹H-NMR′(500 MHz,DMSO-d6): 1.82-1.91 (4H, m), 2.86-2.93 (1H, m), 3.18-3.36 (4H, m), 4.52(2H, s), 6.90-6.93 (2H, m), 7.30 (1H, dt, J=1.6, 8.5 Hz), 7.65 (1H, brs), 8.00 (1H, d, J=7.6 Hz), 8.97 (1H, br), 9.32 (1H, br).

Example 23-1

(1) With the use of the starting compounds 1G, 2B,and similar processesas those of the step (1) and (2) of Example 22-1, tert-butyl3-{2-amino-3-(aminomethyl)-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-pyridinyl}-1-piperidinecarboxylatewas prepared.

(2) To a cold (0° C.) solution of tert-butyl3-{2-amino-3-(aminomethyl)-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-pyridinyl}-1-piperidinecarboxylate(1.9 g, 4.055 mmol) and triethylamine (1.13 mL, 8.109 mmol) intetrahydrofuran (40.0 mL) was added acetic anhydride (0.457 mL, 4.865mmol). The mixture was stirred at 0° C. for 4 hrs. After quenched bywater, the mixture was extracted with ethyl acetate and water. Theseparated organic phase was washed with brine, dried over MgSO₄,filtered and concentrated under reduced pressure. The resulting residue(yellow solid) was purified by recrystallization from ethylacetate/diisopropyl ether to give tert-butyl3-{3-(acetylamino-methyl)-2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-pyridinyl}-1-piperidinecarboxylate(1.13 g, yield; 55%).

(3) The tert-butyl carbamate was treated in a similar manner as that ofthe step (3) of Example 1-1 to giveN-{[2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-(3-piperidinyl)-3-pyridinyl]methyl}acetamidehydrochoride.

Molecular weight: 446.98; Mass spectrometry: 411 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 0.28-0.30 (2H, m), 0.47-0.50 (2H, m), 1.13 (1H, m), 1.85-1.90(4H, m), 1.93 (3H, s), 2.88 (1H, m), 3.06 (1H, m), 3.23 (1H, m), 3.33(1H, m), 3.73 (1H, m), 3.80 (2H, m), 4.32-4.39 (2H, dd, J=5.6, 15.5 Hz),6.58 (1H, d, J=8.5 Hz), 6.65 (1H, d, J=8.2 Hz), 7.09 (1H, s), 7.27 (1H,t, J=8.2 Hz), 7.80 (1H, br), 8.98 (2H, br), 9.29 (1H, br), 13.63 (1H,br).

Example 23-2

(1) To a cold (0° C.) solution of tert-butyl3-{2-amino-3-(aminomethyl)-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-pyridinyl}-1-piperidinecarboxylate(0.300 g, 0.640 mmol), which was obtained in the step (1) of Example23-1, and triethylamine (0.067 mL, 0.704 mmol) in tetrahydrofuran (30mL) was added ethyl chloroformate (0.134 mL, 0.960 mmol) under an argonatmosphere. The mixture was allowed to warm to room temperature, and thestirring was continued for 4 hrs. After quenched by water, the mixturewas extracted with ethyl acetate. The separated organic phase was washedwith brine, dried over MgSO₄, filtered, then concentrated under reducedpressure. The resulting residue was purified by column chromatography onSilica-gel (hexane/ethyl acetate=1/1) to give tert-butyl3-(2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-3-{[(ethoxycarbonyl)amino]methyl}-4-pyridinyl)-1-piperidinecarboxylateas a pale yellow form (0.246 g, yield; 71%).

(2) Then the tert-butyl carbamate was treated in a similar manner asthat of the step (3) of Example 1-1 to give ethyl[2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-(3-piperidinyl)-3-pyridinyl]methylcarbamatehydrochloride.

Molecular weight: 477.01; Mass spectrometry: 441 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 0.28-0.30 (2H, m), 0.48-0.51 (2H, m), 1.17 (3H, t, J=7.1 Hz),1.22 (1H, m), 1.82-1.89 (5H, m), 2.88 (1H, m), 3.03 (1H, m), 3.25 (1H,m), 3.69 (1H, m), 3.84 (2H, m), 4.03 (2H, q, J=6.9 Hz), 4.28 (2H, dd,J=5.4, 15.1 Hz), 6.59 (1H, d, J=8.2Hz), 6.65 (1H, d, J=8.2 Hz), 7.11(1H, s), 7.27 (1H, t, J=8.2 Hz), 7.64 (1H, s), 7.86 (1H, s), 8.80 (1H,s), 9.27 (1H, s).

Example 23-3

(1) To a solution of tert-butyl3-{2-amino-3-aminomethyl}-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-pyridinyl}-1-piperidinecarboxylate(0.150 g, 0.320 mmol), which was obtained in the step (1) of Example23-1, in tetrahydrofuran (10.0 mL) was added cyclohexyl isocyanate(0.044 g, 0.352 mmol). The mixture was stirred at room temperature for1.5 hrs and concentrated under reduced pressure. The resulting residuewas purified by preparative TLC (ethyl acetate) to give tert-butyl3-[2-amino-3[(3-cyclohexyl-ureido)-methyl]-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-pyridinyl]-1-piperidinecarboxylateas a yellow form (0.113 g, yield; 60%).

(2) Then the tert-butyl carbamate was treated in a similar manner asthat of the step (3) of Example 1-1 to giveN-{[2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-(3-piperidinyl)-3-pyridinyl]methyl}-N′-cyclohexylureahydrochloride.

Molecular weight: 530.12; Mass spectrometry: 494 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 0.27-0.28 (2H, m), 0.47-0.49 (2H, m), 1.10-1.14 (4H, m),1.22-1.27 (3H, m), 1.51 (1H, m), 1.62-1.65 (2H, m), 1.73-1.74 (3H, m),1.83-1.89 (5H, m), 2.88 (1H, m), 3.01 (1H, m), 3.72 (1H, m), 3.78-3.85(2H, m), 4.29-4.32 (2H, m), 6.59 (1H, d, J=8.5 Hz), 6.66 (1H, d, J=8.2Hz), 7.05 (1H, s), 7.27 (1H, t, J=8.2 Hz), 8.98 (1H, s), 9.25 (1H, s),10.3 (1H, s), 13.5 (1H, s).

Examples 23-4 to 23-15

According to the similar synthetic procedure of Examples 23-1 to 23-3,compounds shown in Table 6 were prepared.

TABLE 6 Mol in Ex. No Structure weight Mass vitro A549 NMR 23-04

376.89 341 A A (500 MHz, DMSO-d6): 1.80-1.95(4H, m), 2.90-2.95(1H, m),3.13-3.34(2H, m), 4.26(1H, dd, J=5.8, 15.2Hz), 4.34 (1H, dd, J=5.8,15.2Hz), 6.95(1H, t, J=7.3Hz), 6.98(1H, d, J=8.2Hz), 7.22 (1H, s),7.35(1H, t, J=8.2Hz), 7.67(1H, br s), 8.81(1H, br), 8.93(1H, br), 9.26(1H, br), 13.66(1H, br). 23-05

402.93 367 A A (500 MHz, DMSO-d6): 0.78(4H, m), 1.68 (1H, m), 1.88(4H,m), 1.99(1H, m), 3.05 (2H, dd, J=11.4, 23.9Hz), 3.38(2H, m), 3.49(1H,m), 4.34(2H, ddd, J=6.0, 13.5, 44.4Hz), 6.94(1H, t, J=7.6Hz), 7.00(1H,d, J=8.2Hz), 7.23(1h, S), 7.35(1H, t, J=8.2Hz), 7.70(1H, br), #8.90(1H,br), 9.02(1H, br), 9.21(1H, br), 13.7(1H, br). 23-06

390.92 355 B B (500 MHz, DMSO-d6): 1.03(3H, t, J= 7.6Hz), 1.85(2H, d,J=13.3Hz), 1.99 (1H, m), 2.18(2H, q, J=7.6Hz), 3.05 (2H, dd, J=11.4,23.9Hz), 3.38(2H, m), 3.49(1H, m), 4.33(2H, d, J=6.0 Hz), 6.97(2H, t,J=7.9Hz), 7.07(1H, m), 7.37(1H, t, J=7.9Hz), 7.56(1H, br #s), 7.88(1H,br), 8.75(1H, br), 8.96(1H, m), 13.6(1H, br). 23-07

446.98 411 A A (500 MHz, DMSO-d6): 0.28-0.31(2H, m), 0.47-0.50(2H, m),1.18(1H, m), 1.85-1.91(3H, m), 1.92(3H, s), 1.94- 2.00(2H, m),3.04-3.09(2H, m), 3.49- 3.57(2H, m), 3.83(2H, d, J=6.9Hz), 4.35(2H, d,J=5.7Hz), 6.60(1H, d, J= 8.5Hz), 6.68(1H, d, J=8.2Hz), 6.83 #(1H, m),7.27(1H, dd, J=8.2, 8.5Hz), 7.84(1H, br s), 8.96(3H, m), 10.4(1H, br s),13.6(1H, br s). 23-08

491.04 455 A A (300 MHz, DMSO-d6): 0.23-0.35(2H, m), 0.41-0.57(2H, m),1.19(6H, d, J= 6.4Hz), 1.06-1.28(1H, m), 1.70-1.98 (4H, m),2.76-3.15(2H, m), 3.15-3.60 (2H, m), 3.60-3.75(1H, m), 3.75-3.90 (2H,m), 4.15-4.45(2H, m), 4.70-4.90 (1H, m), 6.59(1H, d, J=8.3Hz), 6.66#(1H, d, J=8.3Hz), 7.11(1H, br s), 7.27 (1H, t, J=8.3Hz), 7.61(1H, br),7.80 (1H, br s), 8.65-8.95(1H, m), 9.15- 9.40(1H, m), 10.38(1H, br),23-09

505.06 469 A A (300 MHz, DMSO-d6): 0.22-0.35(2H, m), 0.41-0.56(2H, m),0.88(6H, d, J= 6.8Hz), 1.02-1.25(1H, m), 1.70-2.00 (5H, m),2.75-3.15(2H, m), 3.15-3.65 (2H, m), 3.65-3.97(5H, m), 4.15-4.44 #(2H,m), 6.59(1H, d, J=8.3Hz), 6.67 (1H, d, J=8.3Hz), 7.10(1H, br s), 7.27(1H, t, J=8.3Hz), 7.69(1H, br), 7.92 (1H, br s), 8.94(1H, br), 9.41(1H,br), 10.41(1H, br), 13.68(1H, br). 23-10

519.09 483 A A (300 MHz, DMSO-d6): 0.24-0.35(2H, m), 0.40-0.57(2H, m),0.89(9H, s), 1.06-1.25(1H, m), 1.70-2.00(4H, m), 2.75-3.15(2H, m),3.15-3.50(2H, m), 3.72(2H, s), 3.67-3.90(3H, m), 4.24 (1H, dd, J=5.7,15.5Hz), 4.37(1H, dd, J=5.7, 15.5Hz), 6.58(1H, d, J=8.3Hz), #6.67(1H, d,J=8.3Hz), 7.11(1H, br s), 7.26(1H, t, J=8.3Hz), 7.67(1H, br), 7.91(1H,br s), 8.95(1H, br), 9.39(1H, br), 10.39(1H, br), 13.68(1H, br). 23-11

476.02 440 A A (500 MHz, DMSO-d6): 0.28-0.29(2H, m), 0.47-0.49(2H, m),1.13(1H, m), 1.85-1.90(5H, m), 2.86(6H, s), 3.01 (1H, m), 3.25-3.27(2H,m), 3.78-3.84 (2H, m), 3.96(1H, m), 4.31(2H, d, J= 5.0Hz), 6.59(1H, d,J=7.9Hz), 6.64 (1H, d, J=7.0Hz), 7.04(1H, s), 7.27 #(1H, t, J=7.5Hz),8.00(1H, s), 9.09(2H, s), 10.35(1H, s). 23-12

490.05 454 A A (500 MHz, DMSO-d6): 0.28-0.29(2H, m), 0.47-0.49(2H, m),0.82(3H, t, J= 7.4Hz), 1.13(1H, m), 1.37-1.42(2H, m), 1.84-1.89(4H, m),2.90(1H, m), 3.00-3.01(3H, m), 3.02-3.05(2H, m), #3.78-3.85(3H, m),4.30-4.36(2H, m), 6.58(1H, d, J=8.5Hz), 6.67(1H, d, J= 8.2Hz), 7.04(1H,s), 7.18(1H, s), 7.26 (1H, t, J=8.2Hz), 8.06(2H, s), 9.05(1H, s),9.39(1H, s), 10.39(1H, s), 13.60(1H, s). 23-13

447.97 412 A A (500 MHz, DMSO-d6): 0.28-0.29(2H, q), 0.47-0.49(2H, q),1.14(1H, br s), 1.85-1.89(4H, m), 2.88(1H, m), 3.02 (1H, m),3.25-3.33(2H, m), 3.71(1H, m), 3.78-3.86(2H, m), 4.28-4.34(2H, m),5.95(1H, br), 6.58(1H, d, J=8.2 Hz), 6.67(1H, d, J=8.2Hz), 7.04(1H, s),7.26(1H, t, J=8.2Hz), 8.09(2H, br), #9.04(1H, br), 9.40(1H, br),10.40(1H, br), 13.35(1H, s). 23-14

524.07 488 A A (500 MHz, DMSO-d6): 0.27-0.28(2H, m), 0.47-0.48(2H, m),1.12(1H, m), 1.88- 1.90(4H, m), 2.89-3.09(2H, m), 3.35- 3.38(3H, m),3.70-3.85(2H, m), 4.38- 4.45(2H, m), 6.58(1H, d, J=8.8Hz), 6.66(1H, d,J=8.2Hz), 6.91-7.00(1H, m), 7.10(1H, s), 7.25(3H, t, J=7.6Hz), #7.40(2H,d, J=7.6Hz), 7.82(1H, s), 8.85(1H, s), 9.28(2H, s), 10.37(1H, s). 23-15

540.13 504 A A (500 MHz, DMSO-d6): 0.29-0.31(2H, m), 0.48-0.50(2H, m),1.14(1H, m), 1.81-1.92(4H, m), 2.89(1H, m), 3.08 (1H, m), 3.59-3.60(2H,m), 3.61(1H, m), 3.82(2H, dd, J=6.2, 10.1Hz), 4.85 (2H, dd, J=5.4,15.6Hz), 6.59(1H, d, J=8.5Hz), 6.65(1H, d, J=8.2Hz), 7.12 #(1H, t,J=7.5Hz), 7.14(1H, s), 7.27(1H, t, J=8.5Hz), 7.32(2H, t, J=7.6Hz),7.49(2H, d, J=7.8Hz), 7.70(1H, br), 8.71(2H, br), 9.25(1H, br),10.23(1H,

Example 24-1

To a cold (0° C.) solution of tert-butyl3-{3-[(acetylamino)methyl]-2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]4-pyridinyl}-1-piperidinecarboxylate(1.134 g, 2.220 mmol), which was obtained in the step (1) of Example23-1, and triethylamine (1.871 mL, 13.320 mmol) in tetrahydrofuran (150mL) was added dropwise a solution of triphosgene (0.652 g, 2.220 mmol)in tetrahydrofuran (150 mL) under an argon atmosphere. After stirredovernight, the reaction mixture was quenched by water. The mixture wasextracted with ethyl acetate. The separated organic phase was washedwith brine, dried over MgSO₄, filtered and concentrated under reducedpressure. The resulting residue (yellow solid) was purified by columnchromatography on silica gel (hexane/ethyl acetate=1/1) to givetert-butyl3-{3-acetyl-7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-1,2,3,4-tetrahydropyrido[2,3-d]pyrimidin-5-yl}-1-piperidinecarboxylateas a white form (0.569 g, yield; 48%).

Then the tert-butyl carbamate was treated in a similar manner as that ofthe step (3) of Example 1-1 to give3-acetyl-7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-5-(3-piperidinyl)-3,4-dihydropyrido[2,3-d]pyrimidin-2(1H)-onehydrochloride.

Molecular weight: 472.98; Mass spectrometry: 437 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 0.31-0.34 (2H, m), 0.56-0.59 (2H, m), 1.26 (1H, m), 1.74-1.79(2H, m), 1.81-1.93 (2H, m), 2.47 (H, s), 2.85-2.98 (2H, m), 3.21-3.34(3H, m), 3.81-3.86 (2H, m), 4.93 (2H, s), 6.52 (2H, d, J=8.2 Hz), 7.16(1H, t, J=8.2 Hz), 7.78 (1H, s), 8.79 (1H, br), 9.30 (1H, br), 10.95(1H, s).

Example 24-2

To a cooled (0° C.) solution of tert-butyl3-(2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-3-{[(ethoxycarbonyl)amino]methyl}-4-pyridinyl)-1-piperidinecarboxylate(0.145 g, 0.268 mmol), which was obtained in the step (1) of Example23-2, and triethylamine (0.226 mL, 1.609 mmol) in tetrahydrofuran(30 mL)was added triphosgene (0.079 g, 0.268 mmol) in tetrahydrofuran (10 mL)under an argon atmosphere. The mixture was allowed to warm to roomtemperature, and the stirring was continued for 3 hrs. After quenched bywater, the mixture was extracted with ethyl acetate. The separatedorganic phase was washed with water and brine successively, dried overMgSO₄, filtered, and concentrated under reduced pressure. The resultingresidue was purified by preparative TLC (hexane/ethyl acetate=1/1) togive ethyl5-[1-(tert-butoxycarbonyl)-3-piperidinyl]-7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-1,4-dihydropyrido[2,3-d]pyrimidine-3(2H)-carboxylateas a white solid (0.904 g, yield; 60%).

Then the tert-butyl ester was treated in a similar manner as that of thestep (3) of Example 1-1 to give ethyl7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-5-(3-piperidinyl)-1,4-dihydropyrido[2,3-d]pyrimidine-3(2H)-carboxylatehydrochloride.

Molecular weight: 503.00; Mass spectrometry: 467 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR(500 MHz,DMSO-d6): 0.31-0.34 (2H, m), 0.55-0.59 (2H, m), 1.26-1.28 (1H, br s),1.27 (3H, t, J=7.3 Hz), 1.74-1.85 (3H, m), 2.85-2.95 (2H, m), 3.22-3.25(1H, m), 3.30-3.32 (2H, m), 3.79-3.87 (1H, m), 4.22-4.25 (2H, m), 4.91(2H, s), 6.52 (2H, d, J=8.2 Hz), 7.16 (1H, t, J=8.2 Hz), 7.78 (1H, s),8.91 (1H, br s), 9.29 (1H, br), 10.86 (1H, s).

Example 24-3

To a cooled (0° C.) solution of tert-butyl3-{2-amino-3-({[(cyclohexylamino)-carbonyl]amino}methyl)-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-pyridinyl}-1-piperidinecarboxylate(0.039g, 0.065 mmol), which was obtained in the step (1) of Example 23-3, andtriethylamine (15.0 mL, 0.389 mmol) in tetrahydrofuran (10.0 mL) wasadded dropwise a solution of triphosgene (0.019 g, 0.065 mmol) intetrahydrofuran (5.0 mL) under an argon atmosphere. The mixture wasstirred at 0° C. for 2 hrs. After quenched by water, the mixture wasextracted with ethyl acetate. The separated organic phase was washedwith brine, dried over MgSO₄, filtered and concentrated under reducedpressure. The resulting residue (white solid) was triturated withdiisopropyl ether, collected by filtration, washed with diisopropylether, and dried under reduced pressure to give tert-butyl3-{3-[(cyclohexylamino)carbonyl]-7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-1,2,3,4-tetrahydropyrido[2,3-d]pyrimidin-5-yl}-1-piperidinecarboxylate(0.021 g, yield; 52%).

Then the tert-butyl carbamate was treated in a similar manner as that ofthe step (3) of Example 1-1 to giveN-cyclohexyl-7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-5-(3-piperidinyl)-1,4-dihydropyrido[2,3-d]pyrimidine-3(2H)-carboxamidehydrochloride.

Molecular weight: 556.11; Mass spectrometry: 520 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR′(500 MHz,DMSO-d6): 0.32-0.34 (2H, m), 0.55-0.59 (2H, m), 1.24-1.35 (8H, m), 1.64(1H, m), 1.75-1.78 (2H, m), 1.82-1.84 (4H, m), 1.91-1.93 (2H, m), 2.86(1H, m), 2.96 (1H, m), 3.16 (1H, m), 3.62 (1H, m), 3.63-3.88 (2H, dd,J=6.9, 9.8 Hz), 4.98 (2H, s), 6.52 (2H, d, J=8.2 Hz), 7.16 (1H, t, J=8.5Hz), 7.77 (1H, s), 9.00 (1H, s), 10.82 (1H, s), 11.73 (1H, s).

Example 24-4

To a cold (0° C.) solution of tert-butyl3-{2-amino-3-(aminomethyl)-6-[2-(cyclo-propylmethoxy)-6-hydroxyphenyl]-4-pyridinyl}-1-piperidinecarboxylate(0.150 g, 0.320 mmol), which was obtained in the step (1) of Example23-1, and triethylamine (0.067 mL, 0.480 mmol) in tetrahydrofuran (9 mL)was added methanesulfonyl chloride (0.027 mL, 0.352 mmol) under an argonatmosphere. The stirring was continued at 0° C. for 1 hr. After quenchedby water, the mixture was extracted with ethyl acetate. The separatedorganic phase was washed with brine, dried over MgSO₄, filtered, andconcentrated under reduced pressure. The resulting yellow residue waspurified by recrystallization from diisopropyl ether/dichloromethane togive tert-butyl3-(2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-3-{[(methylsulfonyl)amino]methyl}-4-pyridinyl)-1-piperidinecarboxylateas a yellow solid (0.097 g, yield; 56%).

To a cooled (0° C.) solution of tert-butyl3-(2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-3-{[(methylsulfonyl)amino]methyl}-4-pyridinyl)-1-piperidinecarboxylate(0.050 g, 0.091 mmol) and triethylamine (0.077 mL, 0.549 mmol) intetrahydrofuran (15 mL) was added to a solution of triphosgene (0.027 g,0.091 mmol) in tetrahydrofuran(5 mL) under an argon atmosphere. Themixture was allowed to warm to room temperature, and the stirring wascontinued for 3 hrs under an argon atmosphere. After quenched by water,the mixture was extracted with ethy acetate. The separated organic phasewas washed with brine, dried over MgSO₄, filtered, and concentratedunder reduced pressure. The resulting yellow solid was triturated withdiisopropyl ether and collected by filtration to give tert-butyl3-[7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-3-(methylsulfonyl)-2-oxo-1,2,3,4-tetrahydropyrido[2,3-d]pyrimidin-5-yl]-1-piperidinecarboxylateas a yellow solid (0.028 g, yield; 55%).

Then the tert-butyl carbamate was treated in a similar manner as that ofthe step (3) of Example 1-1 to give7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-3-(methylsulfonyl)-5-(3-piperidinyl)-3,4-dihydropyrido[2,3-d]pyrimidin-2(1H)-onehydrochloride.

Molecular weight: 509.03; Mass spectrometry: 473 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR′(500 MHz,DMSO-d6): 0.30-0.33 (2H, m), 0.54-0.58 (2H, m), 1.26 (1H, m), 1.73-1.78(2H, m), 1.88-1.93 (2H, m), 2.88 (1H, m), 2.99 (1H, m), 3.13 (1H, m),3.30-3.35 (2H, m), 3.50 (3H, s), 3.83 (2H, dd, J=6.9, 7.2 Hz), 4.97 (2H,s), 6.53 (2H, d, J=8.2 Hz), 7.17 (1H, t, J=8.5 Hz), 7.75 (1H, s), 8.67(1H, s), 9.11 (1H, s), 11.02 (1H, s).

Examples 24-5 to 24-10

According to the similar synthetic procedure of Examples 24-1 to 24-4,compounds shown in Table 7 were prepared.

TABLE 7 Mol in Ex. No Structure weight Mass vitro A549 NMR 24-05

517.03 481 A A (300 MHz, DMSO-d6): 0.25-0.38(2H, m), 0.50-0.64(2H, m),1.29(6H, d, J= 6.0Hz), 1.20-1.32(1H, m), 1.60-2.00 (4H, m),2.75-3.05(2H, m), 3.15-3.40 (3H, m), 3.75-3.92(2H, m), 4.89(2H, s),4.90-5.04(1H, m), 6.53(2H, d, J=8.3 Hz), 7.17(1H, t, J=8.3Hz), 7.78(1H,s), #8.92(1H, br), 9.25(1H, br), 10.82(1H, s). 24-06

531.06 495 A A (300 MHz, DMSO-d6): 0.25-0.38(2H, m), 0.51-0.64(2H, m),0.95(6H, d, J= 6.4Hz), 1.17-1.35(1H, m), 1.63-2.05 (5H, m),2.75-3.10(2H, m), 3.10-3.45 (3H, m), 3.75-3.92(2H, m), 3.99(2H, d,J=8.3Hz), 4.89(1H, d, J=15.8Hz), 4.95(1H, d, J=15.8Hz), 6.53(2H, d, J=#8.3Hz), 7.17(1H, t, J=8.3Hz), 7.78 (1H, br), 8.65-9.20(1H, m),9.20-9.75 (1H, m), 10.84(1H, s). 24-07

545.08 509 A A (300 MHz, DMSO-d6): 0.25-0.37(2H, m), 0.52-0.63(2H, m),0.97(9H, s), 1.17-1.35(1H, m), 1.60-2.00(4H, m), 2.75-3.10(2H, m),3.10-3.40(2H, m), 3.76-3.92(3H, m), 3.90(2H, s), 4.89 (1H, d, J=15.8Hz),4.96(1H, d, J=15.8 Hz), 6.53(2H, d, J=8.3Hz), 7.17(1H, t, #J=8.3Hz),7.77(1H, s), 8.65-9.05(1H, m), 9.25-9.60(1H, m), 10.83(1H, s). 24-08

502.02 466 A A (500 MHz, DMSO-d6): 0.33-0.35(2H, m), 0.56-0.58(2H, m),1.26(1H, m), 1.76-1.80(2H, m), 1.90-1.92(2H, m), 2.84-2.91(10H, m),3.14(1H, m), 3.83- 3.85(2H, m), 4.64-4.70(2H, m), 6.51- 6.54(2H, m),7.16(1H, t, J=8.2Hz), 7.80(1H, s), 8.47(1H, s), 8.97(1H, s), #10.68(1H,s). 24-09

502.02 466 (500 MHz, DMSO-d6): 0.32-0.33(2H, m), 0.56-0.58(2H, m),1.10(3H, t, J= 7.3Hz), 1.27(1H, m), 1.72-1.91(4H, m), 2.85-2.95(2H, m),3.16(1H, m), 3.22-3.27(2H, m), 3.31-3.33(2H, m), 3.82-3.86(2H, m),4.99(2H, s), 6.52 (2H, d, J=8.2Hz), 7.17(1H, t, J=8.2 #Hz), 7.78(1H, s),8.96(1H, br), 9.17(1H, br), 10.84(1H, s). 24-10

473.96 438 A A (300 MHz, DMSO-d6): 0.32-0.33(2H, m), 0.56-0.58(2H, m),1.26(1H, m), 1.73-1.95(3H, m), 2.85-2.97(3H, m), 3.19-3.34(4H, m),3.78-3.86(2H, m), 4.98(2H, a), 6.53(2H, d, J=7.9Hz), 7.16(1H, t,J=8.3Hz), 7.46(1H, br), 7.78(1H, s), 8.33(1H, br), 8.67(1H, br),#9.12(1H, br), 10.80(1H, s).

To a suspension of tert-butyl3-[2-amino-3-formyl-6-(2-hydroxyphenyl)4-pyridinyl]-1-piperidinecarboxylate(0.86 g, 2.16 mmol) in ethanol (25 mL) were added diethyl malonate (6.93g, 43.27 mmol) and piperidine (2.14 mL, 21.64 mmol) and, and the mixturewas heated at reflux overnight. The mixture was allowed to cool to roomtemperature, and then diluted with ethanol. The resulting precipitatewas collected by filtration, washed with ethanol and dried under reducedpressure to give ethyl5-[1-(tert-butoxycarbonyl)-3-piperidinyl]-7-(2-hydroxyphenyl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carboxylateas a yellow solid (0.752 g, yield; 71%).

Then the tert-butyl carbamate was treated in a similar manner as that ofthe step (3) of Example 1-1 to give ethyl7-(2-hydroxyphenyl)-2-oxo-5-(3-piperidinyl)-1,2-dihydro-1,8-naphthyridine-3-carboxylatehydrochloride.

Molecular weight: 429.91; Mass spectrometry: 394 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-B; ¹H-NMR (500 MHz,DMSO-d6): 1.34 (3H, t, J=7.1 Hz), 1.89-2.07 (4H, m,), 2.92-3.00 (1H, m),3.40-3.44 (1H, m, 3.77 (1H, br), 4.32 (2H, q, J=7.1 Hz), 6.97-7.01 (2H,m), 7.41 (1H, t, J=7.25 Hz), 8.01 (1H, s), 8.21 (1H, d, J=7.25 Hz), 8.62(1H, s), 8.83 (1H, br), 9.18 (1H, br), 12.69 (1H, s), 12.96 (1H, br s).

Example 25-2

(1) To a stirred solution of tert-butyl3-[2-amino-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-3-(hydroxymethyl)-4-pyridinyl]-1-piperidinecarboxylate(1.100 g, 1.863 mmol), which was obtained in the step (2) of Example17-1, in dichloromethane (50 mL) was added manganese dioxide (3.240 g,37.261 mmol). The mixture was stirred at room temperature for 3 hrs andfiltered on Celite®. The filtrate was concentrated under reducedpressure to give tert-butyl3-(2-amino-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-3-formyl-4-pyridinyl)-1-piperidinecarboxylate(0.978 g, yield; 89%).

(2) To a stirred solution of tert-butyl3-(2-amino-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-3-formyl-4-pyridinyl)-1-piperidinecarboxylate(0.500 g, 5 0.851 mmol), triethyl-2-fluoro-2-phosphonoacetate (0.350 mL,1.701 mmol) and anhydrous lithium chloride (0.072 g, 1.701 mmol) inacetonitrile (20 mL) was added 1,8-diazabicyclo[5,4,0]undec-7-ene (0.250mL, 1.701 mmol). The mixture was stirred at room temperature for 30 minand concentrated under reduced pressure. The residue was extracted withethyl acetate and water. The separated organic phase was washed withbrine, dried over Na₂SO₄, filtered, and concentrated under reducedpressure. The resulting residue (liquid) was purified by columnchromatography on silica gel (hexane/ethyl acetate=2/1-1/1) to givetert-butyl3-(2-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-6-fluoro-7-oxo-7,8-dihydro-1,8-naphthyridin-4-yl)-1-piperidinecarboxylateas a form (0.261 g, 49%).

Then the tert-butyl carbamate was treated in a similar manner as that ofthe step (2) of Example 21-1 to give7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-3-fluoro-5-(3-piperidinyl)-1,8-naphthyridin-2(1H)-onehydrochloride.

Molecular weight: 445.93; Mass spectrometry: 410 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 0.32 (2H, m), 0.55 (2H, m), 1.25 (1H, m), 1.94 (4H, m), 2.95(2H, m), 3.65 (1H, br), 6.58 (2H, d, J=8.2 Hz), 7.23 (1H, t, J=8.2 Hz),7.97 (1H, s), 8.15 (1H, d, J=11.4 Hz), 8.64 (1H, br), 9.01 (1H, br),11.66 (1H, br), 13.02 (1H, d, J=5.4 Hz).

Example 25-3

(1) With the use of the starting compound 1G and tert-butyl 4-formylpiperidine-1-carboxylic acid, tert-butyl4-(2-amino-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl)-3-formyl-4-pyridinyl)-1-piperidinecarboxylatewas prepared in a similar manner as that of the step (1) of Example25-2. To a solution of tert-butyl4-(2-amino-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-3-formyl-4-pyridinyl)-1-piperidinecarboxylate(0.400 g, 0.681 mmol) in ethyl alcohol (5.0 mL) was added ethyl malonatemonoamine (1.780 g, 13.612 mmol) and piperidine (0.580 g, 6.806 mmol).The mixture was refluxed 12 hrs. The reaction mixture was extracted withethyl acetate and water. The separated organic phase was washed withNaHCO₃ and brine, dried over Na₂SO₄, filtered, and concentrated underreduced pressure. The resulting residue (liquid) was purified by columnchromatography on silica gel (hexane/ethyl acetate=1/1 and ethylacetate100%) to give tert-butyl4-(6-(aminocarbonyl)-2-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-7-oxo-7,8-dihydro-1,8-naphthyridin-4-yl)-1-piperidinecarboxylateas a yellow form. (0.455 g, yield; quant.)

(2) Then the tert-butyl carbamate was treated in a similar manner asthat of the step (2) of Example 21-1 to give7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-5-(4-piperidinyl)-1,2-dihydro-1,8-naphthyridine-3-carboxamidehydrochloride.

Molecular weight: 470.96; Mass spectrometry: 435 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-B; ¹H-NMR(500 MHz,DMSO-d6): 0.32 (2H, d, J=4.7 Hz), 0.57 (2H, d, J=7.9 Hz), 1.32-1.35 (1H,m), 1.91-2.01 (5H, m), 3.23-3.26 (3H, m), 3.70-3.80 (2H, m), 3.90 (2H,d, J=6.9 Hz), 6.59 (2H, dd, J=1.7, 8.3 Hz), 7.26 (1H, t, J=8.2 Hz), 7.87(1H, d, J=3.5 Hz), 8.01 (1H, s), 8.70 (1H, d, J=12.3 Hz), 8.87 (1H, d,J=10.7 Hz), 8.99 (1H, d, J=3.4 Hz), 9.06 (1H, s), 13.05 (1H, s).

Example 25-4

(1) To a solution of ethyl (diphenoxyphosphoryl)acetate (1.090 g, 3.403mmol) in THF (15 mL) were added 1,8-diazabicyclo[5,4,0]undec-7-ene(0.520 g, 3.403 mmol) and NaI (0.510 g, 3.403 mmol) followed by asolution of tert-butyl3-(2-amino-6-(2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]-phenyl}-3-formyl-4-pyridinyl)-1-piperidinecarboxylate(1.000 g, 1.701 mmol), obtained in the step (1) of Example 25-2, in THF(5 mL). The mixture was stirred at 0° C. for 1.5 hrs and quenched withsaturated aqueous NaHCO₃ solution. The mixture was extracted with ethylacetate, and the extract was washed with brine, dried over Na₂SO₄,filtered, and concentrated under reduced pressure. The residue waspurified by column chromatography on silica gel (hexane/ethylacetate=3:2 to 1:2) to give tert-butyl3-(2-{2-(cyclo-propylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-7-oxo-7,8-dihydro-1,8-naphthyridin-4-yl)-1-piperidinecarboxylateas an amorphous solid (0.276 g, 27%).

(2) To a stirred solution of tert-butyl3-(2-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-7-oxo-7,8-dihydro-1,8-naphthyridin-4-yl)-1-piperidine-carboxylate(0.260 g, 0.425 mmol) in 1,4-dioxane (2 mL) was added 4 N HCl in dioxane(2 mL). The mixture was stirred at room temperature overnight, anddiluted with ethyl acetate. The resulting precipitate was collected byfiltration, washed with ethanol, and dried under reduced pressure togive7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-5-(3-piperidinyl)-1,8-naphthyridin-2(1H)-onehydrochloride as a yellow solid (0.146 g, 80%).

Molecular weight: 427.93; Mass spectrometry: 392 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 0.34 (2H, m), 0.58 (2H, m), 1.28 (1H, m), 1.91 (4H, m), 2.89(1H, br d, J=6.6 Hz), 3.00 (1H, dd, J=10.4, 12.3 Hz), 3.37 (2H, dd,J=12.3, 12.6 Hz), 6.57 (2H, d, J=8.2 Hz), 6.67 (1H, d, J=9.8 Hz), 7.23(1H, t, J=8.2 Hz), 7.99 (1H, s), 8.21 (1H, d, J=9.8 Hz), 9.02 (1H, br),9.14 (1H, br), 12.45 (1H,br s).

Examples 25-5 to 25-8

According to the similar synthetic procedure of Examples 25-1 to 25-4,compounds shown in Table 8 were prepared.

TABLE 8 Mol in Ex. No Structure weight Mass vitro A549 NMR 25-05

472.93 437 A B (500 MHz, DMSO-d6): 0.33(2H, dd, J= 4.7, 9.1Hz), 0.56(2H,m), 1.25(1H, m), 1.94(4H, m), 2.90(1H, br), 3.00(1H, t, J= 12.3Hz),3.40(2H, br d, J=12.3Hz), 6.59(2H, d, J=8.2Hz), 7.27(1H, t, J= 8.2Hz),8.03(1H, s), 9.02(1H, s), 11.57 (1H, s), 13.32(1H, br). 25-06

452.94 417 A A (500 MHz, DMSO-d6): 0.32(2H, dd, J= 4.7, 9.1Hz), 0.57(2H,m), 1.25(1H, m), 1.93(4H, m), 2.89(1H, br d, J=7.3Hz), 2.97(1H, dd,J=11.4, 11.7Hz), 3.39 (2H, t, J=14.5Hz), 6.59(2H, d, J=8.5 Hz), 7.26(1H,t, J=8.5Hz), 8.00(1H, s), 8.81(1H, br), 9.01(1H, s), 9.26(1H, br),#11.66(1H, s), 13.13(1H, s). 25-07

568.10 532 A A (500 MHz, DMSO-d6): 0.32(2H, m), 0.57 (2H, m), 1.25(1H,m), 1.95(4H, m), 2.93 (1H, br d, J=11.0Hz), 3.06(1H, dd, J= 11.0,12.0Hz), 6.58(2H, dd, J=2.5, 8.2 Hz), 7.25(1H, t, J=8.2Hz), 7.64(2H, t,J=7.6Hz), 7.74(1H, t, J=7.6Hz), 8.04 (3H, m), 8.85(1H, br), 9.01(1H, s),9.35 #(1H, br), 11.47(1H, br), 12.93(1H, s). 25-08

470.96 435 A A (500 MHz, DMSO-d6): 0.33(2H, m), 0.58 (2H, m), 1.28(1H,m), 1.90(4H, m), 2.91 (1H, d, J=11.3Hz), 3.06(1H, dd, J= 11.3, 12.3Hz),6.59(2H, d, J=8.2Hz), 7.26(1H, t, J=8.2Hz), 7.90(1H, d, J= 3.5Hz),8.06(1H, s), 8.81(1H, br), 9.00 (1H, d, J=3.5Hz), 9.02(1H, s), 9.31#(1H, br), 11.80(1H, br), 13.07(1H, s).

Example 26-1

(1) To a cold (0° C.) suspension of ethyl5-[1-(tert-butoxycarbonyl)-3-piperidinyl]-7-(2-hydroxyphenyl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carboxylate(0.050 g, 0.101 mmol) in THF (2.0 mL) under an argon atmosphere wasadded LiBH₄ (0.004 g, 0.20 mmol). The mixture was allowed to warm toroom temperature, and the stirring was continued for 3 hrs. Theresulting mixture was quenched with water, and partitioned between ethylacetate and a saturated ammonium chloride solution. The separatedorganic phase was washed with water and brine, dried over Na₂SO₄,filtered and concentrated under reduced pressure. The residue waspurified by column chromatography on silica gel (hexane: ethyl acetate,3:2) to give Ethyl5-[1-(tert-butoxycarbonyl)-3-piperidinyl]-7-(2-hydroxyphenyl)-2-oxo-1,2,3,4-tetrahydro-1,8-naphthyridine-3-carboxylate.(0.027 g, yield; 54%)

(2) Ethyl5-[1-(tert-butoxycarbonyl)-3-piperidinyl]-7-(2-hydroxyphenyl)-2-oxo-1,2,3,4-tetrahydro-1,8-naphthyridine-3-carboxylate(0.024 g, 0.048 mmol) was treated under acidic conditions in a similarmanner as described in Example 1-1 to give ethyl7-(2-hydroxyphenyl)-2-oxo-5-(3-piperidinyl)-1,2,3,4-tetrahydro-1,8-naphthyridine-3-carboxylate.(0.019 g, yield; 91%)

Molecular weight: 431.92; Mass spectrometry: 396 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR′(500 MHz,DMSO-d6): 1.16-1.21 (3H, m), 1.83-1.95 (4H, m), 2.87-2.94 (1H, m),3.18-3.37 (6H, m), 4.12-4.17 (2H, m), 6.90-6.93 (2H, m), 7.29 (1H, t,J=7.88 Hz), 7.73 (1H, s), 8.03 (1H, d, J=7.88 Hz), 8.81-8.90 (1H, m),9.23-9.30 (1H, m), 11.33 (1H, s).

Example 26-2

To a cold (0° C.) solution of4-(6-(aminocarbonyl)-2-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-7-oxo-7,8-dihydro-1,8-naphthyridin-4-yl)-1-piperidine-carboxylate(0.200 g, 0.305 mmol), which was obtained in the step (1) of Example25-3, in methyl alcohol (5.0 mL) under an argon atmosphere was addedNaBH₄ (0.010 g, 0.367 mmol). The mixture was allowed to warm to roomtemperature, and the stirring was continued for 12 hrs. The reactionmixture was quenched with water, and extracted with ethyl acetate. Theseparated organic phase was washed with brine, dried over Na₂SO₄,filtered, and concentrated under reduced pressure to give tert-butyl4-(6-(aminocarbonyl)-2-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)-oxy]phenyl}-7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-4-yl)-1-piperidinecarboxylate.(0.133 g, yield; 67%)

To a stirred solution of tert-butyl4-(6-(aminocarbonyl)-2-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-4-yl)-1-piperidinecarboxylate(0.120 g, 0.187 mmol) in 1,4-dioxane (3 mL) was added 4N HCl in dioxane(1 mL). The mixture was stirred at room temperature for 2 hrs. Theresulted precipitate was collected by filtration, washed withacetonitrile, and dried under reduced pressure to give7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-5-(4-piperidinyl)-1,2,3,4-tetrahydro-1,8-naphthyridine-3-carboxamidehydrochloride. (0.080 g, yield 90%)

Molecular weight: 472.98; Mass spectrometry: 437 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 0.33 (2H, d, J=4.4 Hz), 0.60 (2H, d, J=6.3 Hz), 1.33-1.39 (1H,m), 1.77-1.91 (5H, m), 3.06-3.10 (2H, m), 3.20 (3H, d, J=8.2 Hz), 3.86(2H, dd, J=2.2, 7.1 Hz), 6.53 (2H, dd, J=8.2, 7.0 Hz), 7.16 (1H, t,J=8.2 Hz), 7.20 (1H, s), 7.56 (1H, s), 7.84 (1H, s), 11.02 (1H, s).

Examples 26-3 to 26-6

According to the similar synthetic procedure of Examples 26-1 to 26-2,compounds shown in Table 9 were prepared.

TABLE 9 Mol in Ex. No Structure weight Mass vitro A549 NMR 26-03

502.02 466 A A (300 MHz, DMSO-d6): 0.31-0.36(2H, m), 0.56-0.60(2H, m),1.17(3H, t, J= 7.2Hz), 1.26(1H, m), 1.82-1.90(4H, m), 2.84-2.89(2H, m),3.23-3.39(6H, m), 3.80-3.89(2H, m), 4.16(2H, q, J= 7.2Hz), 6.52(2H, d,J=8.3Hz), 7.16 (1H, t, J=8.3Hz), 7.85(1H, s), 9.17(1H, #br), 9.39(1H,br), 11.17(1H, s). 26-04

502.02 466 A A (500 MHz, DMSO-d6): 0.30-0.33(2H, m), 0.56-0.59(2H, m),1.17(3H, t, J= 7.2Hz), 1.36(1H, m), 1.86-1.92(4H, m), 3.06-3.09(2H, m),3.18-3.23(2H, m), 3.30-3.38(3H, m), 3.80(1H, m), 3.85-3.87(2H, d,J=7.0Hz), 4.12(2H, q, J=7.2Hz), 6.51(1H, d, J=8.2Hz), #6.53(1H, d,J=8.2Hz), 7.16(1H, t, J= 8.2Hz), 7.83(1H, s), 8.96(1H, br), 9.03 (1H,br), 11.16(1H, s). 26-05

472.98 437 A A (500 MHz, DMSO-d6): 0.36(2H, m), 0.60 (2H, m), 1.30(1H,m), 1.73-1.94(4H, m), 2.94(2H, m), 6.52, 6.53(2H, d × 2, J=8.2Hz),7.16(1H, t, J=8.2Hz), 7.22, 7.23(1H, br × 2), 7.54, 7.59(1H, br s × 2),7.85, 7.87(1H, s × 2), 8.86 1H, br), #9.15(1H, br), 11.04, 11.04(1H, s ×2). 26-06

490.00 454 A A (500 MHz, DMSO-d6): 0.93(3H, t, J= 7.3Hz), 1.17(3H, td,J=3.5, 7.2Hz), 1.74-1.79(3H, m), 1.85-1.91(3H, m), 2.87-2.89(2H, m),3.34-3.40(3H, m), 3.80-3.85(1H, m), 3.93-3.97(2H, m), 4.14(2H, q,J=7.2Hz), 6.52(1H, d, J= 8.2Hz), 6.57(1H, d, J=8.5Hz), 7.17 #(1H, t,J=8.2Hz), 7.65(1J, s), 8.86(1H, br), 9.15(1H, br), 11.13(1H, s).

A solution of ethyl7-(2-hydroxyphenyl)-2-oxo-5-(3-piperidinyl)-1,2,3,4-tetrahydro-1,8-naphthyridine-3-carboxylatehydrochloride (0.012 g, 0.023 mmol), which was obtained in the step(2)of Example 26-1, in 2.5N HCl (3.0 mL) was heated at reflux for 4 hrs.After being cooled to room temperature, the mixture was concentratedunder reduced pressure. The residue was washed with acetonitrilee anddried under reduced pressure to give7-(2-hydroxyphenyl)-5-(3-piperidinyl)-3,4-dihydro-1,8-naphthyridin-2(1H)-onehydrochloride. (0.007 g, yield; 70%)

Molecular weight: 359.86; Mass spectrometry: 324 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 1.83-1.94 (4H, m), 2.57-2.64 (2H, m), 2.89-3.01 (3H, m),3.19-3.37 (4H, m), 6.88-6.93 (2H, m), 7.28 (1H, t, J=8.2 Hz), 7.69 (1H,s), 8.01 (1H, d, J=7.9 Hz), 8.90 (1H, br), 9.16 (1H, br), 10.99 (1H, s).

Examples 27-2 to 27-5

In similar manners as described in Example 27-1 above, compounds inExamples 27-2 to 27-5 were synthesized.

According to the similar synthetic procedure of Example 27-1, compoundsshown in Table 10 were prepared.

TABLE 10 Mol in Ex. No Structure weight Mass vitro A549 NMR 27-02

417.94 382 A A (500 MHz, DMSO-d6): 0.94(3H, t, J= 7.5Hz), 1.72-1.80(2H,m), 1.87- 1.89(4H, m), 2.59-2.61(2H, m), 2.88- 2.91(2H, m),2.99-3.02(2H, m), 3.26-3.33(2H, m), 3.43(1H, m), 3.95 (2H, m), 6.54(1H,d, J=8.2Hz), 6.58 (1H, d, J=7.9Hz), 7.18(1H, t, J=7.4 #Hz), 7.64(1H, s),9.26(1H, br), 10.84 (1H, s). 27-03

429.95 394 A A (300 MHz, DMSO-d6): 0.32- 0.37(2H, m), 0.56-0.62(2H, m),1.29 (1H, m), 1.75-1.83(4H, m), 2.57- 2.61(2H, m), 2.88(1H, m),2.96-3.01 (2H, m), 3.33-3.36(4H, m), 3.82- 3.87(2H, m), 6.52(2H, d,J=7.9Hz), 7.15(1H, t, J=7.9Hz), 7.83(1H, s), 9.07(1H, br), 10.81(1H, s).27-04

429.95 394 A A (500 MHz, DMSO-d6): 0.31-0.34(2H, m), 0.57-0.60(2H, m),1.33(1H, m), 1.81-1.85(2H, m), 1.90-1.93(2H, m), 2.57(2H, t, J=7.2Hz),2.99(2H, t, J=7.2Hz), 3.06-3.08(2H, m), 3.18 (1H, m), 3.38-3.40(2H, m),3.85(2H, d, J=6.9Hz), 6.52(2H, d, J=7.9Hz), #7.15(1H, t, J=8.2Hz),7.80(1H, s), 8.52(1H, br), 8.77(1H, br), 10.80(1H, s). 27-05

417.94 382 A A (500 MHz, DMSO-d6): 0.93(3H, t, J= 7.3Hz), 1.80-1.84(2H,m), 2.57- 2.59(2H, m), 2.99-3.08(4H, m), 3.12 (1H, m), 3.36-3.39(2H, m),3.97(2H, t, J=7.7Hz), 6.52(1H, d, J=8.2Hz), 6.57(1H, d, J=8.5Hz),7.18(1H, t, J= 8.2Hz), 7.59(1H, s), 9.03(2H, br), 10.80(1H, s).

To a stirred solution of7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-5-(3-piperidinyl)-3,4-dihydro-1,8-naphthyridin-2(1H)-one(0.029 g, 0.074 mmol), which was obtained in the Example 27-3, in TBF(1.5 mL) was added dropwise Vitride® (70% toluene solution, 0.500 mL).The mixture was stirred at 65° C. for 1 hr. After cooled to roomtemperature, the mixture was quenched with saturated aqueous potassiumsodium tartrate solution. The mixture was extracted with ethyl acetateand water. The separated organic phase was washed with brine, dried overNa₂SO₄, filtered, and concentrated under reduced pressure. The resultingresidue was purified by preparative TLC (chloroform/methanol=4/1) togive3-(cyclopropylmethoxy)-2-[4-(3-piperidinyl)-5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl]phenol.(0.003 g, yield; 11%)

Molecular weight: 379.51; Mass spectrometry: 380 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,CDCl3): 0.42 (2H, m), 0.68 (2H, m), 1.35 (1H, m), 1.91 (1H, d, J=12.0Hz), 1.98 (2H, m), 2.63 (1H, dt, J=2.8, 12. Hz), 2.77 (3H, m), 2.86 (1H,m), 3.11 (2H, dd, J=2.3, 10.5 Hz), 3.40 (2H, dt, J=2.8, 5.4 Hz), 3.83(2H, d, J=6.9 Hz), 4.75 (1H, br s), 6.38 (1H, dd, J=1.3, 8.2 Hz), 6.59(1H, dd, J=1.3, 8.2 Hz), 7.09 (1H, t, J=8.2 Hz), 7.78 (1H, s), 14.69(1H, br).

Example 29-01

(1) To a stirred solution of tert-butyl3-(2-amino-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-3-formyl-4-pyridinyl)-1-piperidinecarboxylate(0.100 g, 0.170 mmol), which was obtained in the step (1) of Example25-2, and ethyl nitroacetate (0.060 mL, 0.510 mmol) in ethanol (5 mL)was added piperidine (0.050 mL, 0.510 mmol). The mixture was stirred at75° C. for 18 hrs, and concentrated under reduced pressure. The residuewas extracted with ethyl acetate and water. The separated organic phasewas washed with brine, dried over Na₂SO₄, filtered, and concentratedunder reduced pressure. The resulting residue (liquid) was purified bycolumn chromatography on silica gel (hexane/ethyl acetate=3/1-3/2) togive tert-butyl3-(2-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-6-nitro-7-oxo-7,8-dihydro-1,8-naphthyridin-4-yl)-1-piperidinecarboxylate(0.105 g, yield; 94%).

(2) To a stirred mixture of tert-butyl3-(2-{2-(cyclopropylmethoxy)-6-[(4-methoxy-benzyl)oxy]phenyl}-6-nitro-7-oxo-7,8-dihydro-1,8-naphthyridin-4-yl)-1-piperidine-carboxylate (0.085 g, 0.129 mmol), iron powder (0.300 g),ethanol (4.5 mL) and water (0.5 mL) was added ammonium chloride (0.100g). The mixture was stirred at 85° C. for 30 min. After cooled to roomtemperature, the mixture was filtered through Celite®. The filtrate wasconcentrated under reduced pressure. The residue was extracted withethyl acetate and water. The separated organic phase was washed withbrine, dried over Na₂SO₄, filtered, and concentrated under reducedpressure to give tert-butyl3-(6-amino-2-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]-phenyl}-7-oxo-7,8-dihydro-1,8-naphthyridin-4-yl)-1-piperidinecarboxylate(0.068 g, yield; 84%).

(3) Then benzyl moiety and tert-butyl carbamate were removed in asimilar manner as described in the step (2) of Example 25-4, to give3-amino-7-[2-(cyclo-propylmethoxy)-6-hydroxyphenyl]-5-(3-piperidinyl)-1,8-naphthyridin-2(1H)-onehydrochloride.

Molecular weight: 442.95; Mass spectrometry: 407 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 0.35 (2H, m), 0.62 (2H, m), 1.32 (1H, m), 1.73 (1H, ddd,J=4.1, 8.2, 8.2 Hz), 1.93 (3H, m), 2.90 (1H, dd, J=11.7, 12.3 Hz), 3.11(1H, dd, J=11.0, 11.7 Hz), 6.54 (2H, d, J=8.2Hz), 6.93 (1H, s), 7.17(1H, t, J=8.2 Hz), 7.99 (1H, s), 9.10 (1H, br), 9.28 (1H, br), 12.55(1H, s).

Example 29-2

To a stirred solution of tert-butyl3-(6-amino-2-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-7-oxo-7,8-dihydro-1,8-naphthyridin-4-yl)-1-piperidine-carboxylate(0.068 g, 0.109 mmol), which was obtained in the step (2) of Example29-1, and triethyl amine (0.05 mL, 0.327 mmol) in methylene chloride (3mL) were added benzoyl chloride (0.01 mL, 0.109 mmol) and4-dimethylaminopyridine (0.001 g, 0.011 mmol). The mixture was stirredat room temperature for 19 hrs, and poured into water. The mixture wasextracted with ethyl acetate. The separated organic phase was washedwith brine, dried over Na₂SO₄, filtered, and concentrated under reduced.The resulting residue was purified by column chromatography on silicagel (hexane/ethyl acetate=5/1-3/2) to give tert-butyl3-{2-{2-(cyclopropylmethoxy)-6-[(4-methoxy-benzyloxy]phenyl}-7-oxo-[(1-phenyl-methanoyl)-amino]-7,8-dihydro-1,8-naphthyridin-4-yl)-1-piperidinecarboxylate(0.022 g, yield; 28%).

Then benzyl moiety and tert-butyl carbamate were removed in a similarmanner as described in the step (2) of Example 25-4 to giveN-[7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-5-(3-piperidinyl)-1,2-dihydro-1,8-naphthyridin-3-yl]benzamidehydrochloride.

Molecular weight: 547.06; Mass spectrometry: 511 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-B; ¹H-NMR (500 MHz,DMSO-d6): 0.34 (2H, m), 0.59 (2H, m), 1.29 (1H, m), 1.80-2.08 (4H, m),2.96 (1H, br), 3.18 (1H, dd, J=11.4, 11.7 Hz), 6.59 (2H, d, J=8.2 Hz),7.23 (1H, t, J=8.2 Hz), 7.61 (2H, t, J=7.9 Hz), 7.68 (1H, m), 8.01 (2H,m), 8.04 (1H, s), 8.80 (1H, br), 9.03 (2H, s), 9.61 (1H, s), 13.13 (1H,s).

Example 29-3

In a similar manner as that of Example 29-2,N-[7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-5-(3-piperidinyl)-1,2-dihydro-1,8-naphthyridin-3-yl]acetamidehydrochloride was prepared.

Molecular weight: 484.99; Mass spectrometry: 449 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; (500 MHz,DMSO-d6): 0.33 (2H, m), 0.59 (2H, m), 1.28 (1H, m), 1.76 (1H, m), 1.89(1H, m), 1.98 (1H, m), 2.92 (1H, dd, J=11.4, 12.0 Hz), 3.17 (1H, ddJ=11.0, 12.0 Hz), 6.57 (2H, d, J=8.5 Hz), 7.21 (1H, t, J=8.5 Hz), 7.99(1H, s), 8.78 (1H, br), 8.94 (1H, s), 9.02 (1H, br), 9.69 (1H, s), 11.92(1H, br), 12.93 (1H, s).

EXAMPLE 30-1

(1) To a stirred solution of ethyl5-[1-(tert-butoxycarbonyl)-3-piperidinyl]-7-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carboxylate(0.276 g, 0.403 mmol), which was obtained in a similar manner as that ofthe step (1) of Example 29-1 using diethyl malonate instead of ethylnitroacetate, in tetrahydrofuran (4 mL) and water (1 mL) was addedlithium hydroxide monohydrate (0.015 g, 0.605 mmol). The mixture wasstirred at room temperature for 18 hrs and acidified (pH 3-4) withaqueous 1N hydrochloric acid solution. The mixture was extracted withethyl acetate. The separated organic phase was washed with brine, driedover Na₂SO₄, filtered, and concentrated under reduced pressure to give5-[1-(tert-butoxycarbonyl)-3-piperidinyl]-7-{2-(cyclopropyl-methoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carboxylicacid (0.259 g, yield; 98%).

(2) Then benzyl moiety and tert-butyl carbamate were removed in asimilar manner as described in the step (2) of Example 25-4 to give7-(2-cyclopropylmethoxy-6-hydroxy-phenyl)-2-oxo-5-piperidin-3-yl-1,2-dihydro-[1,8]naphthyridine-3-carboxylicacid hydrochloride.

Molecular weight: 471.94; Mass spectrometry: 436 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-B; ′(500 MHz,DMSO-d6): 0.30-0.33 (2H, m), 0.54-0.55 (2H, m), 1.23 (1H, m), 1.92-1.98(4H, m), 2.91 (1H, m), 3.04 (1H, m), 3.39-3.41 (3H, m), 3.86 -3.89 (3H,m), 6.61 (2H, d, J=8.5 Hz), 7.27 (1H, t, J=8.2 Hz), 8.07 (1H, s), 8.72(1H, s), 9.24 (1H, s), 13.74 (1H, s), 14.43 (1H, s).

Example 31-1

(1) To a stirred solution of5-[1-(tert-butoxycarbonyl)-3-piperidinyl]-7-{2-(cyclo-propylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carboxylicacid (0.200 g, 0.305 mmol), which was obtained in the step (1) ofExample 30-1, and 1-hydroxybenzotriazole hydrate (0.062 g, 0.457 mmol)in dimethylformamide (4 mL) was added1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.070 g,0.366 mmol). The mixture was stirred at room temperature for 18 hrs, andthen extracted with ethyl acetate and water. The separated organic phasewas washed with brine, dried over Na₂SO₄, filtered, and concentratedunder reduced pressure. The resulting residue was purified by columnchromatography on silica gel (hexane/ethyl acetate=1/1) to givetert-butyl3-{2-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-7-oxo-6-[(propylamino)-carbonyl]-7,8-dihydro-1,8-naphthyridin-4-yl}-1-piperidinecarboxylate(0.129 g, yield; 61%).

(2) Then benzyl moiety and tert-butyl carbamate were removed in asimilar manner as described in the step (2) of Example 25-4 to give7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-5-(3-piperidinyl)-N-propyl-1,2-dihydro-1,8-naphthyridine-3-carboxamide.

Molecular weight: 513.04; Mass spectrometry: 477 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 0.33 (2H, m), 0.57 (2H, m), 0.94 (3H, t, J=7.3 Hz), 1.27 (1H,m), 1.58 (2H, tq, J=7.3, 7.3 Hz), 1.92 (4H, m), 2.90 (1H, m), 3.06 (1H,t, J=12.3 Hz), 6.59 (2H, d, J 8.2 Hz), 7.26 (1H, t, J=8.2 Hz), 8.07 (1H,s), 9.01 (1H, s), 9.69 (1H, t, J=6.0 Hz), 11.80 (1H, s).

Example 31-2

To a stirred solution of methanesulfonamide (0.026 g, 0.268 mmol) andtriethylamine (0.030 mL, 0.229 mmol) in toluene (2 mL) was addedtrimethylsilyl chloride (0.020 mL, 0.152 mmol). The mixture was stirredat 90° C. for 1 hr, and then concentrated under reduced pressure. To theresidue diluted with dichloromethane (2 mL) were added triethylamine(0.020 mL, 0.152 mmol), 4,4-dimethylaminopyridine (0.002 g, 0.015 mmol),benzotriazole-1-yloxy-tris(dimethylamino)phosphoniumhexafluoro-phosphate (0.040 g, 0.091 mmol) and5-[1-(tert-butoxycarbonyl)-3-piperidinyl]-7-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carboxylicacid (0.050 g, 0.076 mmol) successively. The mixture was stirred at roomtemperature for 6 hrs. The mixture was extracted with ethyl acetate andwater. The separated organic phase was washed with brine, dried overNa₂SO₄, filtered, and concentrated under reduced pressure. The resultingresidue was purified by preparative TLC (hexane/ethyl acetate=1/1×2) togive tert-butyl3-(2-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-6-{[(methylsulfonyl)-amino]carbonyl}-7-oxo-7,8-dihydro-1,8-naphthyridin-4-yl)-1-piperidinecarboxylate(0.024 g, yield; 43%).

Then the benzyl moiety and tert-butyl carbamate were removed in asimilar manner as described in the step (2) of Example 25-4 to giveN-{[7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-5-(3-piperidinyl)-1,2-dihydro-1,8-naphthyridin-3-yl]carbonyl}methanesulfonamidehydrochloride.

Molecular weight: 549.05; Mass spectrometry: 513 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-B; ¹H-NMR (500 MHz,DMSO-d6): 0.33 (2H, m), 0.55 (2H, m), 1.25 (1H, m), 1.95 (4H, m), 2.92(1H, br), 3.06 (1H, t, J=12.0 Hz), 3.40 (2H, t, J=11.7 Hz), 3.45 (3H,s), 6.61 (2H, d, J=8.2 Hz), 7.28 (1H, t, J=8.2 Hz), 8.09 (1H, s), 8.74(1H, br), 9.08 (1H, s), 11.62 (1H, br), 12.72 (1H, br), 13.56 (1H, br).

Example 31-3 to 31-27

According to the similar synthetic procedure of Example 31-1 to 31-2,compounds shown in Table 11 were prepared.

TABLE 11 in Ex. Mol vi- No Structure weight Mass tro A549 NMR 31-03

484.99 449 A A (500 MHz, DMSO-d6): 0.34(2H, m), 0.57(2H, m), 1.27(1H,m), 1.92(4H, m), 2.91(3H, d, J=4.7Hz), 3.05(1H, t, J= 12.3Hz), 6.59(2H,d, J=8.2Hz), 7.26 (1H, t, J=8.2Hz), 8.06(1H, s), 9.00(1H, s), 9.53(1H,q, J=4.7Hz), 11.79(1H, s). 31-04

499.01 463 A B (500 MHz, DMSO-d6): 0.34(2H, m), 0.57(2H, m), 1.27(1H,m), 1.94(4H, m), 2.92(3H, s), 3.00(3H, s), 6.58(2H, d, J= 8.2Hz),7.24(1H, t, J=8.2Hz), 8.00 (1H, s), 8.19(1H, s), 8.58(1H, br), 9.10 (1H,br), 11.84(1H, br), 12.72(1H, s). 31-05

525.05 489 A B (500 MHz, DMSO-d6): 0.33(2H, m), 0.57(2H, m), 1.25(1H,m), 1.89(9H, m), 2.90(1H, m), 2.98(1H, dd, J=11.4, 12.6 Hz), 6.58(2H, d,J=8.2Hz), 7.24(1H, t, J=8.2Hz), 8.00(1H, s), 8.22(1H, d, J= 2.5Hz),8.46(1H, br), 9.05(1H, br), 11.82(1H, br), 12.74(1H, s). 31-06

561.09 525 A A (500 MHz, DMSO-d6): 0.33(2H, m), 0.57(2H, m), 1.27(1H,m), 1.92(4H, m), 2.91(1H, m), 3.06(1H, t, J=12.3Hz), 4.61(2H, d,J=6.0Hz), 6.59(2H, d, J= 8.5Hz), 7.28(2H, m), 7.37(4H, m), 8.08(1H, s),9.05(1H, s), 10.07(1H, t, J=6.0Hz), 11.80(1H, s). 31-07

529.04 493 A A (500 MHz, DMSO-d6): 0.33(2H, m), 0.57(2H, m), 1.27(1H,m), 1.92(4H, m), 2.91(1H, m), 3.07(1H, t, J=12.3Hz), 3.50(2H, m),3.55(2H, m), 6.60(2H, d, J=8.2Hz), 7.26(1H, t, J=8.2Hz), 8.07(1H, s),9.02(1H, s), 9.78(1H, m), 11.80(1H, s). 31-08

541.05 505 A B (500 MHz, DMSO-d6): 0.33(2H, m), 0.57(2H, m), 1.26(1H,m), 1.93(4H, m), 2.90(1H, m), 2.98(1H, dd, J=11.3, 12.0 Hz), 3.73(1H,br), 3.88(2H, m), 6.58 (2H, d, J=8.2Hz), 7.24(1H, t, J=8.2 Hz), 8.00(1H,s), 8.63(1H, br), 9.06 (1H, br d, J=10.7Hz), 11.85(1H, br), 12.73(1H,s). 31-09

576.53 504 A C (500 MHz, DMSO-d6): 0.33(2H, m), 0.57(2H, m), 1.26(1H,m), 1.94(4H, m), 2.90(1H, m), 2.98(1H, dd, J=10.7, 11.4 #Hz), 3.18(4H,br s), 3.88(5H, m), 6.58(2H, d, J=8.2Hz), 7.24(1H, t, J=8.2Hz), 8.00(1H,s), 8.38(1H, s), 9.03-9.21 (4H, br), 11.81(1H, br), 12.78(1H, s). 31-10

513.04 477 A A (500 MHz, DMSO-d6): 0.34(2H, m), 0.57(2H, m), 1.23(6H, d,J=6.6Hz), 1.88-1.99(4H, m), 2.91(1H, m), 3.07 (1H, dd, J=11.0, 12.0Hz),4.11(1H, dq, J=6.6, 7.6Hz), 6.59(2H, d, J=8.2Hz), 7.26(1H, t, J=8.2Hz),8.07(1H, s), 8.72 (1H, br), 9.01(1H, s), 9.59(1H, d, J=7.6 #Hz),11.80(1H, br), 13.13(1H, s). 31-11

578.54 506 A A (500 MHz, DMSO-d6): 0.33(2H, m), 0.56(2H, m), 1.26(1H,m), 1.88-1.99 (4H, m), 2.85(6H, s), 3.11(1H, dd, J= 11.0, 12.3Hz),3.75(2H, t, J=5.7Hz), 6.59(2H, d, J=8.2Hz), 7.27(1H, t, J= 8.2Hz),8.07(1H, s), 8.83(1H, br), 9.01 (1H, s), 9.14(1H, br), 9.66(1H, br),9.83 #(1H, t, J=6.0Hz), 11.75(1H, s), 13.18 (1H, s). 31-12

587.12 551 A A (500 MHz, DMSO-d6): 0.33(2H, m), 0.57(2H, m), 1.26(1H,m), 1.88-1.99 (4H, m), 2.92(3H, dd, J=5.4, 10.7Hz), 3.07(1H, t,J=12.3Hz), 4.74(1H, ddd, J=6.9, 6.9, 12.6Hz), 6.59(2H, d, J= 8.2Hz),7.19(2H, m), 7.28(3H, m), 8.06(1H, s), 9.03(1H, s), 9.91(1H, d, J=#6.9Hz), 11.76(1H, s), 13.09(1H, br). 31-13

637.18 601 A A (500 MHz, DMSO-d6): 0.32(2H, m), 0.56(2H, m), 1.25(1H,m), 1.88-1.97 (4H, m), 2.90(1H, m), 3.05(1H, t, J= 12.3Hz), 6.33(1H, d,J=7.9Hz), 6.60 (2H, d, J=8.2Hz), 7.28(3H, m), 7.38 (8H, m), 8.04(1H, s),9.03(1H, s), 10.07 (1H, d, J=7.9Hz), 11.69(1H, s), 13.20 (1H, br). 31-14

575.11 539 A A (500 MHz, DMSO-d6): 0.33(2H, m), 0.56(2H, m), 1.26(1H,m), 1.53(3H, d, J=6.9Hz), 1.86-1.98(4H, m), 2.91 (1H, m), 3.06(1H, m),5.19(1H, dq, J= 6.9, 7.3Hz), 6.59(2H, d, J=8.2Hz), 7.28(2H, m), 7.39(4H,m), 8.06(1H, d, J=2.9Hz), 8.71(1H, br t, J=11.7Hz), #9.01(1H, d,J=2.9Hz), 9.09(1H, br t, J= 11.7Hz), 10.11(1H, dd, J=5.7, 7.6Hz),11.78(1H, br), 13.16(1H, s). 31-15

575.11 539 A A (500 MHz, DMSO-d6): 0.33(2H, m), 0.56(2H, m), 1.26(1H,m), 1.53(3H, d, J=6.9Hz), 1.86-1.98(4H, m), 2.91(1H, m), 3.06(1H, m),5.19(1H, dq, J=6.9, 7.3Hz), 6.59, 6.59(2H, d × 2, J=8.2 Hz), 7.28(2H,m), 7.39(4H, m), 8.06 (1H, d, J=2.9Hz), 8.73(1H, br t, J= #11.7Hz),9.00(1H, d, J=2.9Hz), 9.16 (1H, br t, J=11.4Hz), 10.11(1H, dd, J= 5.7,7.6Hz), 11.78(1H, br), 13.16(1H, s). 31-16

543.02 507 A B (500 MHz, DMSO-d6): 0.34(2H, m), 0.57(2H, m), 1.28(1H,m), 1.44(3H, d, J=7.3Hz), 1.88-1.99(4H, m), 2.92(1H, m), 3.07(1H, dd,J=11.4, 12.0Hz), 4.53 (1H, m), 6.59, 6.61(2H, d × 2, J=8.2 Hz), 7.27(1H,t, J=8.2Hz), 8.08(1H, s), 8.79(1H, br), 9.02(1H, s), 9.28(1H, br),#10.07, 10.08(1H, d × 2, J=6.9Hz), 13.17(1H, s). 31-17

575.11 539 A B (500 MHz, DMSO-d6): 0.33(2H, m), 0.56(2H, m), 1.26(1H,m), 1.88-1.99 (4H, m), 2.88(2H, t, J=7.3Hz), 3.07 (1H, dd, J=11.7,12.0Hz), 3.63(2H, dt, J=6.3, 7.3Hz), 6.59(2H, d, J=8.2Hz), 7.21-7.33(6H,m), 8.05(1H, s), 8.69 (1H, br), 8.99(1H, br d, J=5.1Hz), 9.01 #(1H, s),9.73(1H, t, J=6.3Hz), 11.74 (1H, br s), 13.11(1H, s). 31-18

553.11 517 A A (500 MHz, CDCl3): 0.41-0.47(2H, m), 0.65-0.77(2H, m),0.77-0.92(1H, m), 1.32-1.42(1H, m), 1.78-1.84(1H, m), 1.84-1.97(2H, m),2.15-2.24(1H, m), 2.72(1H, dd, J=2.8, 12.0Hz), 2.80- 2.88(m, 1H),3.16-3.26(1H, m), 3.34- 3.46(2H, m), 3.91(2H, d, J=6.9Hz), #5.95(2H, brs), 6.42(1H, dd, J=1.0, 8.2 Hz), 6.73(1H, dd, J=1.0, 8.2Hz), 7.23-7.26(1H, m), 8.63(1H, s), 8.69(1H, s), 15.00(1H, br). 31-19

567.13 531 A C (500 MHz, DMSO-d6): 0.33(2H, m), 0.56(2H, m), 1.02(2H,m), 1.14-1.26 (4H, m), 1.53(1H, br), 1.64(1H, br d, J= 11.4Hz), 1.72(4H,m), 1.93(4H, m), 2.92(1H, br), 3.07(1H, t, J=12.0Hz), 6.59(2H, d,J=8.2Hz), 7.26(1H, t, J= 8.2Hz), 8.05(1H, s), 8.90(1H, br), 9.00 #(1H,br), 9.01(1H, s), 9.73(1H, d, J= 11.7Hz), 11.74(1H, s), 13.12(1H, br).31-20

515.01 479 A B (500 MHz, DMSO-d6): 0.33(2H, m), 0.56(2H, m), 1.27(1H,m), 1.93(4H, m), 2.91(1H, m), 3.07(1H, t, J=12.0Hz), 3.39(2H, br),3.44(2H, dd, J=5.7, 6.0 Hz), 3.56(2H, dd, J=5.4, 5.7Hz), 6.59 (2H, d,J=8.2Hz), 7.26(1H, t, J=8.2 Hz), 8.07(1H, s), 9.02(1H, s), 9.79(1H, d,J=5.4Hz), 11.82(1H, s), 13.13(1H, br). 31-21

511.03 475 A A (500 MHz, DMSO-d6): 0.34(2H, m), 0.58(2H, m), 0.79(2H,m), 1.26(1H, m), 1.91(4H, m), 2.93(2H, m), 3.07(1H, t, J=12.0Hz),6.60(2H, d, J=8.2Hz), 7.26(1H, t, J=8.2Hz), 8.06(1H, s), 8.99(1H, s),9.66(1H, d, J=4.4Hz), 11.76(1H, s), 13.13(1H, br). 31-22

525.05 489 A A (500 MHz, DMSO-d6): 0.33(2H, m), 0.56(2H, m), 1.26(1H,m), 1.74(2H, m), 1.87-2.05(6H, m), 2.91(1H, m), 3.05 (1H, t, J=12.3Hz),4.46(1H, m), 6.59 (2H, d, J=8.2Hz), 7.26(1H, t, J=8.2Hz), 8.06(1H, s),8.99(1H, s), 9.84(1H, d, J= 7.6Hz), 11.79(1H, s), 13.13(1H, br). 31-23

539.08 503 A A (500 MHz, DMSO-d6): 0.34(2H, m), 0.56(2H, m), 1.26(1H,m), 1.50(2H, m), 1.62(2H, m), 1.71(2H, m), 1.88-1.99 (6H, m), 2.92(1H,m), 3.06(1H, t, J= 12.0Hz), 6.59(2H, d, J=8.2Hz), 7.26 (1H, t, J=8.2Hz),8.06(1H, s), 9.01(1H, s), 9.72(1H, d, J=6.9Hz), 11.77(1H, s), #13.12(1H,br). 31-24

567.13 531 A A (500 MHz, DMSO-d6): 0.34(2H, m), 0.56(2H, m), 1.26(1H,m), 1.59(10H, m), 1.87-1.99(6H, m), 2.90(1H, m), 3.06(1H, t, J=11.7Hz),4.07(1H, m), 6.59(2H, d, J=8.5Hz), 7.26(1H, t, J= 8.5Hz), 8.06(1H, s),9.01(1H, s), 9.78 (1H, d, J=7.9Hz), 11.76(1H, s), 13.12 (1H, br). 31-25

541.10 505 A A (500 MHz, DMSO-d6): 0.34(2H, m), 0.56(2H, m), 0.90(6H, t,J=8.2Hz), 1.27(1H, m), 1.51(2H, m), 1.61(2H, m), 1.88-1.99(4H, m),2.92(1H, m), 3.07 (1H, t, J=12.0Hz), 6.60(2H, d, J=8.2 Hz), 7.26(1H, t,J=8.2Hz), 8.07(1H, s), 8.67(1H, br), 8.98(1H, br), 9.03(1H, s),#9.54(1H, d, J=8.8Hz), 11.78(1H, s), 13.12(1H, br). 31-26

569.15 533 A A (500 MHz, DMSO-d6): 0.34(2H, m), 0.56(2H, m), 0.90(6H, t,J=7.3Hz), 1.25-1.54(9H, m), 1.88-1.99(4H, m), 2.92(1H, m), 3.06(1H, t,J=12.3Hz), 4.07(1H, m), 6.59(2H, d, J=8.2Hz), 7.26(1H, t, J=8.2Hz),8.07(1H, s), 8.69(1H, br), 9.00(1H, br), 9.03(1H, s), #9.51(1H, d,J=9.1Hz), 11.78(1H, s), 13.11(1H, br). 31-27

552.99 517 A A (500 MHz, DMSO-d6): 0.34(2H, m), 0.57(2H, m), 1.27(1H,m), 1.88-1.99 (4H, m), 2.92(1H, m), 3.06(1H, t, J= 12.3Hz), 4.28(2H, m),6.60(2H, d, J= 8.2Hz), 7.27(1H, t, J=8.2Hz), 8.08 (1H, s), 9.07(1H, s),10.12(1H, d, J=6.6 Hz), 11.76(1H, s), 13.26(1H, br).

Example 32-1

With the use of the starting compound 1G, 2B, and other materials,tert-butyl3-(2-amino-3-cyano-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-4-pyridinyl)-1-piperidinecarboxylatewas prepared in a similar manner as that of the step (1) of Example 1-1.

To a solution of tert-butyl3-(2-amino-3-cyano-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-4-pyridinyl)-1-piperidinecarboxylate(0.580 g, 0.999 mmol) in triethyl orthoformate (1.2 mL) was addedammonium sulfate (0.004 g, 0.030 mmol), and the mixture was stirred at150° C. for 2 hrs. After cooled to room temperature, EtOH (1.5 mL) and asolution of NH₃ in EtOH (8.6 N, 0.5 mL) were added successively. Themixture was stirred at room temperature overnight and concentrated underreduced pressure. The residue was triturated with hexane and isopropylether, and dried under reduced pressure to give tert-butyl3-(2-{[(1E)-aminomethylidene]amino}-3-cyano-6-{2-(cyclopropyl-methoxy)-6-[(4-methoxy-benzyl)oxy]phenyl}-4-pyridinyl)-1-piperidinecarboxylateas a brown solid (0.543 g, yield; 89%).

To a solution of tert-butyl3-(2-{[(1E)-aminomethylidene]amino}-3-cyano-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-4-pyridinyl)-1-piperidine-carboxylate(0.180 g, 0.289 mmol) in MeOH (5 mL) and toluene (5 mL) was addedtrifluoroacetic acid (0.01 mL). The mixture was allowed to stand for 20days and concentrated under reduced pressure. Purification by columnchromatography on silica gel (CH₂Cl₂/MeOH 50:1 to 20:1) gave tert-butyl3-(4-amino-7-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]-phenyl}pyrido[2,3-d]pyrimidin-5-yl)-1-piperidinecarboxylateas a colorless oil (0.026 g, yield; 15%).

Then the benzyl moiety and tert-butyl carbamate were removed in asimilar manner as described in the step (2) of Example 25-4 to give2-[4-amino-5-(3-piperidinyl)pyrido[2,3-d]pyrimidin-7-yl]-3-(cyclopropylmethoxy)phenolhydrochloride as a yellow powder (0.004 g, 24%).

Molecular weight: 391.48; Mass spectrometry: 392 (M+H)⁺; In vitroactivity grade: C; Cellular activity grade: (A549)-B; ¹H-NMR (500 MHz,CDCl3): 0.41-0.47 (2H, m), 0.65-0.77 (2H, m), 0.77-0.92 (1H, m),1.32-1.42 (1H, m), 1.78-1.84 (1H, m), 1.84-1.97 (2H, m), 2.15-2.24 (1H,m), 2.72 (1H, dd, J=2.8, 12.0 Hz), 2.80-2.88 (m, 1H), 3.16-3.26 (1H, m),3.34-3.46 (2H, m), 3.91 (2H, d, J=6.9 Hz), 5.95 (2H, br s), 6.42 (1H,dd, J=1.0, 8.2 Hz), 6.73 (1H, dd, J=1.0, 8.2 Hz), 7.23-7.26 (1H, m),8.63 (1H, s), 8.69 (1H, s), 15.00 (1H, br).

Example 33-1

To a solution of oxalyl chloride (3.63 g, 28.6 mmol) in acetonitrile(5.00 mL) at room temperature was added DMF (5.00 mL). After 10 minutes,a solution of 2-(4-methoxyphenylmethoxy)acetophenone (3.33 g, 13.0mmol)(starting compound ID) in DMF (50 mL) was added, and the stirringwas continued overnight. To the reaction mixture were adde malononitrile(1.03 g, 15.6 mmol) and Et₃N (7.25 mL, 52.0 mmol) successively, and thestirring was continued overnight. The reaction mixture was poured intowater, and then extracted with ether. The separated organic phase waswashed with brine, and dried over MgSO₄, filtetred and concentratedunder reduced pressure. The residue was purified by columnchromatography on silica gel (CHCl₃) to give2-((2Z)-3-(dimethylamino)-3-{2-[(4-methoxybenzyl)-oxy]phenyl}2-propenylidene)malononitrile(1.27 g yield; 27%).

To a cold (−78° C.) solution of2-((2Z)-3-(dimethylamino)-3-{2-[(4methoxy-benzyl)oxy]phenyl}-2-propenylidene)malononitrile( 1.26 g, 3.51 mmol) in MeOH was added liquid NH₃, and the mixture washeated at 120° C. in a sealed tube overnight. After cooled to roomtemperature, the reaction mixture was poured into water and theresulting mixture was extracted with ethyl acetate. The separatedorganic phase was washed with brine, and dried over MgSO₄, filtered, andconcentrated under reduced pressure. The crude material was purified bycolumn chromatography on silica gel (hexane:ethyl acetate, 70:30) togive 2-amino-6-({2-[(4-methoxybenzyl)oxy]phenyl}nicotinonitrile (0.050g, yield; 4.3%).

A mixture of 2-amino-6-({2-[(4-methoxybenzyl)oxy]phenyl}nicotinonitrile(0.050 g, 0.151 mmol), trifluoroacetic acid (3.00 mL), anisole (0.50 mL)and water (0.50 mL) was stirred at room temperature overnight. Thereaction mixture was diluted with toluene, and concentrated underreduced pressure. The residue was dissolved in THF, and then hexane wasadded to give precipitates. The precipitates were collected byfiltration and washed with hexane, and then dried under reduced pressureto give 2-amino-6-(2-hydroxyphenyl)nicotinonitrile. (0.026 g, yield;82%)

Molecular weight: 211.23; Mass spectrometry: 212 (M+H)⁺; In vitroactivity grade: B; Cellular activity grade: (A549)-B; ¹H-NMR (500 MHz,DMSO-d6): 6.87-6.92 (2H, m), 7.31-7.39 (4H, m), 7.94 (1H, d, J=8.3 Hz),8.01 (1H, d, J=8.3 Hz), 13.38 (1H, br s).

Example 34-1

(1) To a solution of 2-(benzyloxy)acetophenone (5.66 g, 25mmol)(starting compound 1A) in toluene (40 mL) were added carbondisulfide (6.0 mL, 99.8 mmol) and methyl iodide (10 mL, 245 mmol). Tothe mixture, sodium hydride (60% suspension, 2.00 g, 50 mmol) andN,N-dimethylacetamide (10 mL) were added. The resulting mixture wasstirred at room temperature for 1 hr, and refluxed for 2 hrs. Aftercooled to room temperature, the reaction mixture was partitioned betweenCH₂Cl₂ and water. The organic phase was separated and washed with water,dried, and concentrated under reduced pressure. The residual solid wastriturated with diisopropyl ether to give1-[2-(benzyl)phenyl]-3,3-bis(methylthio)-2-propene-1-one as a lightyellow solid (5.17 g, yield; 63%).

(2) To a suspension of cyanoacetamide (1.27 g, 15.1 mmol) in 2-propanol(50 mL) was added sodium hydride (60% suspension, 0.605 g, 15.1 mmol).The mixture was stirred at room temperature for 10 min. To this mixture,1-[2-(benzyl)phenyl]-3,3-bis(methylthio)-2-propene-1-one (5.00 g, 15.1mmol) was added in one portion. The resulting mixture was refluxed for 4hrs. After being cooled to room temperature, the mixture was dilutedwith 1N HCl. The resulting precipitates were collected by filtration andwashed with EtOH to give6-[2-(benzyloxy)phenyl]-4-(methylthio)-2-oxo-1,2-dihydro-nicotinonitrileas a yellow crystalline solid (4.43 g, yield; 84%).

(3) To a cold (0° C.) solution of6-[2-(benzyloxy)phenyl]-4-(methylthio)-2-oxo-1,2-dihydro-nicotinonitrile(4.00 g, 11.5 mmol) in CH₂Cl₂ was added m-chloroperbenzoic acid (mCPBA)(69%, 3.45 g, 13.8 mmol) in one portion. The reaction mixture wasstirred at 0° C. to room temperature for 1 hr, quenched with saturatedNaHCO₃ solution (80 mL) including Na₂S₂O₃.5H₂O (5 g), and diluted withCH₂Cl₂. The separated organic phase was washed with saturated NaHCO₃solution, dried over Na₂SO₄, filtered, and concentrated under reducedpressure. The residual solid was triturated with diisopropyl ether togive6-[2-(benzyloxy)phenyl]-4-(methylsulfinyl)-2-oxo-1,2-dihydro-nicotinonitrileas a yellow solid (4.00 g, yield; 96%).

(4) A mixture of6-[2-(benzyloxy)phenyl]-4-(methylsulfinyl)-2-oxo-1,2-dihydro-3-pyridinecarbonitrile(2.97 g, 8.16 mmol), bromoacetamide (1.27 g, 9.21 mmol), and K₂CO₃ (1.43g, 10.4 mmol) in DMF (30 mL) was stirred at 60° C. for 1.5 hrs. Thereaction mixture was poured into water, and the resulting precipitateswere collected by filtration. The solid obtained was washed with acetoneto give2-{[6-[2-(benzyloxy)phenyl]-3-cyano-4-(methylsulfinyl)-2-pyridinyl]oxy}acetamideas a solid (2.09 g, yield; 61%).

(5) A mixture of2-{[6-[2-(benzyloxy)phenyl]-3-cyano-4-(methylsulfinyl)-2-pyridinyl]oxy}acetamide(300 mg, 0.71 mmol) and morpholine (1.0 mL) was stirred at 130° C. for 1hr, and then water was added. The resulting precipitates were collectedby filtration and dried under reduced pressure to give2-{[6-[2-(benzyl-oxy)phenyl]-3-cyano-4-(4-morpholinyl)-2-pyridinyl]oxy}acetamideas a yellow solid (304 mg, yield; 96%).

(6) To a solution of2-{[6-[2-(benzyloxy)phenyl]-3-cyano-4-(4-morpholinyl)-2-pyridinyl]oxy}acetamide(318 mg, 0.72 mmol) in DMF (1 mL) was added K₂CO₃ (198 mg, 1.43 mmol),and the mixture was stirred at 130° C. for 24 hrs. The mixture waspartitioned between ethyl acetate and water. The organic phase waswashed with brine, dried over Na₂SO₄, and concentrated under reducedpressure. Purification by column chromatography on silica gel(hexane/ethyl acetate=3/1 to 2/1) followed by trituration withdiisopropyl ether gave2-amino-6-[2-(benzyloxy)phenyl]-4-(4-morpholinyl)nicotinonitrile as awhite solid (141 mg, yield; 51%).

(7) Then the benzyl moiety was removed in a same manner as described inthe step (2) of Example 1-1. The residue was triturated with Et₂O anddried under reduced pressure to give2-amino-6-(2-hydroxyphenyl)-4-(4-morpholinyl)nicotinonitrile as a solid(68 mg, yield; 76%).

Molecular weight: 296.33; Mass spectrometry: 297 (M+H)⁺; In vitroactivity grade: B; Cellular activity grade: (A549)-C; ¹H-NMR (300 MHz,DMSO-d6): 3.44-3.54 (4H, m), 3.68-3.78 (4H, m), 6.77-6.90 (3H, m), 7.08(2H, br s), 7.25-7.35 (1H, m), 7.92-8.00 (1H, m), 13.77 (1H, s).

Example 34-2

A mixture of2-{[6-[2-(benzyloxy)phenyl]-3-cyano-4-(methylsulfinyl)-2-pyridinyl]-oxy}acetamide(500 mg, 1.19 mmol), which was obtained in the step (4) of Example 34-1,and isonipecotamide (228 mg, 1.78 mmol) in DMF (2 mL) was stirred at130° C. for 5 hrs. After cooled to room temperature, the mixture waspartitioned between ethyl acetate and water. The organic phase waswashed with brine, dried over Na₂SO₄ and concentrated under reducedpressure to give1-{2-(2-amino-2-oxoethoxy)-6-[2-(benzyloxy)phenyl]-3-cyano-4-pyridinyl}-4-piperidinecarboxamideas a solid (485 mg, yield; 84%).

A mixture of1-{2-(2-amino-2-oxoethoxy)-6-[2-(benzyloxy)phenyl]-3-cyano-4-pyridinyl}-4-piperidinecarboxamide(485 mg, 1.00 mmol) and K₂CO₃ (280 mg, 2.03 mmol) in DMF (2.5 mL) wasstirred at 130° C. for 28 hrs. After cooled to room temperature, themixture was diluted with water, and the resulting precipitates werecollected by filtration. The precipitates were washed with EtOH andCH₂Cl₂ to give1-{2-amino-6-[2-(benzyloxy)phenyl]-3-cyano-4-pyridinyl}-4-piperidinecarboxamideas a solid (119 mg, yield; 28%).

Then the benzyl moiety was removed in a similar manner as that of thestep (2) of Example 1-1 to give1-[2-amino-3-cyano-6-(2-hydroxyphenyl)-4-pyridinyl]4-piperidinecarboxamideas a solid (19 mg, yield; 36%).

Molecular weight: 337.38; Mass spectrometry: 338 (M+H)⁺; In vitroactivity grade: B; Cellular activity grade: (A549)-C; ¹H-NMR (300 MHz,DMSO-d6): 1.58-1.77 (2H, m), 1.77-1.90 (2H, m), 2.30-2.46 (1H, m),2.98-3.15 (2H, m), 3.97 (2H, d, J=12.8 Hz), 6.73-6.90 (4H, m), 7.10 (2H,s), 7.22-7.36 (2H, m), 7.94 (1H, d, J=7.2 Hz), 13.85 (1H, s).

Example 34-3

To a solution of 1-[(benzyloxy)carbonyl]-3-piperidinecarboxylic acid(3.00 g, 11.4 mmol) and Et₃N (1.56 mL, 11.4 mmol) in t-BuOH (30 mL) wasadded diphenyl phosphoryl azide(DPPA) (2.46 mL, 11.4 mmol), and themixture was refluxed for 21 hrs. After cooled to room temperature, themixture was concentrated under reduced pressure, and the residue waspartitioned between ethyl acetate and a saturated aqueous NaHCO₃solution. The organic phase was washed with a saturated aqueous NaHCO₃solution and brine, dried over Na₂SO₄, and concentrated under reducedpressure. Purification by column chromatography on silica gel(hexane/ethyl acetate=4/1 to 3/1) gave benzyl3-[(tert-butoxycarbonyl)amino]-1-piperidinecarboxylate as a solid (2.22g, yield; 58%).

A mixture of benzyl3-[(tert-butoxycarbonyl)amino]-1-piperidinecarboxylate (2.25 g, 6.73mmol) and 10% Pd/C (0.229 g) in EtOH (20 mL) was stirred at roomtemperature under a hydrogen atmosphere (1 atm) for 14 hrs. The catalystwas removed by filtration through Celite®, and the filtrate wasconcentrated under reduced pressure to give tert-butyl3-piperidinylcarbamate as a solid (1.22 g, yield; 91%).

The resulting tert-butyl 3-piperidinylcarbamate and2-{[6-[2-(benzyloxy)phenyl]-3-cyano-4-(methylsulfinyl)-2-pyridinyl]oxy}acetamidewere used as starting materials to prepare2-amino-4-(3-amino-1-piperidinyl)-6-(2-hydroxyphenyl)nicotinonitrilehydrochloride in a similar manner as described in Example 34-1.

Molecular weight: 345.83; Mass spectrometry: 310 (M+H)⁺; In vitroactivity grade: B; Cellular activity grade: (A549)-B; ¹H-NMR (500 MHz,DMSO-d6): 1.59-1.76 (2H, m), 1.82-1.95 (1H, m), 2.02-2.14(1H, m),3.27-3.50 (3H, m), 3.90-4.05 (1H, m), 4.08-4.22 (1H, m), 6.80 (1H, s),6.92 (1H, t, J=7.6 Hz), 6.96-7.06 (1H, m), 7.37 (1H, t, J=6.9 Hz), 7.79(3H, br s), 8.39 (3H, br s), 13.25 (1H, br s).

Example 34-4

A mixture of2-{[6-[2-(benzyloxy)phenyl]-3-cyano-4-(methylsulfinyl)-2-pyridinyl]-oxy}acetamide(500 mg, 1.19 mol), which was obtained in the step (4) of Example 34-1,and tert-butyl 3-amino-1-piperidinecarboxylate (555 mg, 2.77 mmol) inDMF (0.6 mL) was stirred at 130° C. for 24 hrs. After cooled to roomtemprature, the 10 mixture was partitioned between ethyl acetate andwater. The organic phase was washed with water and brine, dried overNa₂SO₄, filtered, and concentrated under reduced pressure. Purificationby silica gel column chromatography (hexane/ethyl acetate=1/1) gavetert-butyl3-({2-(carbamoylmethoxy)6-[2-(benzyloxy)phenyl]-3-cyano-4-pyridinylamino)-1-piperidinecarboxylateas a foam (360 mg, yield; 64%).

With the use of the resulting compound,2-amino-6-(2-hydroxyphenyl)-4-(3-piperidinylamino)nicotinonitrilehydrochloride was prepared in a similar manner as described in Example34-2.

Molecular weight: 345.83; Mass spectrometry: 310 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (300 MHz,DMSO-d6): 1.68-2.06 (4H, m), 2.66-2.96 (2H, m), 3.17-3.38 (2H, m), 4.23(1H, br s), 6.73 (1H, s), 6.92 (1H, t, J=7.5 Hz), 7.01 (1H, d, J=7.5Hz), 7.37 (1H, t, J=7.5 Hz), 7.55 (3H, br), 7.78 (1H, d, J=7.5 Hz), 9.20(2H, br s), 13.20 (1H, br).

Example 34-5 to 34-19

According to the similar synthetic procedure of Example 34-1 to 34-4,compounds shown in Table 12 were prepared.

TABLE 12 Mol in Ex. No Structure weight Mass vitro A549 NMR 34-05

294.36 295 D (300 MHz, DMSO-d6): 1.64(6H, br s), 3.48(4H, br s),6.76(1H, s), 6.78-6.89 (2H, m), 6.98(1H, br s), 7.28(1H, t, J= 7.2Hz),7.93(1H, d, J=7.2Hz), 13.86 (1H, br s). 34-06

397.44 338 C C (300 MHz, DMSO-d6): 1.49-1.71(2H, m), 1.71-1.87(1H, m),1.91(3H, s), 1.87-2.02(1H, m), 2.37-2.54(1H, m), 2.94-3.08(1H, m),3.09(1H, dd, J= 11.3, 12.8Hz), 3.88-4.04(2H, m), 6.76- 6.92(4H, m),7.01(2H, s), 7.24-7.33 (1H, m), 7.34(1H, br s), 7.95(1H, d, J= #7.2Hz),11.92(1H, br). 34-07

345.83 310 A B (500 MHz, DMSO-d6): 1.90(2H, br s), 1.96(2H, br s),2.98(2H, br s), 3.31(2H, br s), 4.07(1H, br s), 6.67-6.75(1H, m),6.90-6.99(1H, m), 7.04(1H, br s), 7.30- 7.47(1H, m), 7.63(1H, br s),7.81(3H, br), 8.84(1H, br s), 9.09(1H, br s), 12.94 (1H, br). 34-08

345.83 310 B B (500 MHz, DMSO-d6): 2.20(2H, br s), 3.24(2H, br s),3.36(2H, br s), 3.91(2H, br s), 4.09(2H, br s), 6.69(1H, s), 6.94 (1H,t, J=7.6Hz), 7.03(1H, d, J=7.9 Hz), 7.37(1H, t, J=7.9Hz), 7.68(1H, d,J=7.3Hz), 7.84(2H, br), 9.36(2H, br s), 13.15(1H, br). 34-09

331.81 296 A B (500 MHz, DMSO-d6): 3.26(4H, br s), 3.86(4H, br s),6.86(1H, s), 6.92(1H, t, J=7.5Hz), 6.93-7.03(1H, m), 7.36 (1H, t,J=7.5Hz), 7.70(1H, br), 7.82 (1H, d, J=7.5Hz), 9.57(2H, br). 34-10

305.77 270 A A (500 MHz, DMSO-d6): 2.92-3.07(2H, m), 3.71(2H, br s),6.71(1H, d, J=2.8 Hz), 6.94(1H, t, J=7.6Hz), 7.03-7.11 (1H, m), 7.38(1H,t, J=7.6Hz), 7.71 (1H, br s), 7.95(3H, br), 8.13(3H, br s), 12.98(1H,br). 34-11

319.80 284 A B (500 MHz, DMSO-d6): 1.81-1.95(2H, m), 2.79-2.91(2H, m),3.45-3.63(2H, m), 6.67(1H, s), 6.92-6.98(1H, m), 7.10(1H, d, J=8.2Hz),7.35-7.43(1H, m), 7.63(1H, d, J=7.6Hz), 7.85-8.55 (6H, m), 12.80(1H,br). 34-12

319.80 284 A A (500 MHz, DMSO-d6): 2.55-2.61(3H, m), 3.08(2H, br s),3.77(2H, br s), 6.73 (1H, s), 6.94(1H, t, J=7.6Hz), 7.02- 7.11(1H, m),7.38(1H, t, J=7.6Hz), 7.74(1H, br), 7.96(2H, br), 9.06(2H, br),12.99(1H, br). 34-13

395.90 360 A A (500 MHz, DMSO-d6): 3.11(2H, br s), 3.77-3.93(2H, m),4.15-4.23(2H, m), 6.76(1H, s), 6.90-6.98(1H, m), 7.07 (1H, d, J=8.2Hz),7.33-7.48(4H, m), 7.55-7.64(2H, m), 7.66-7.79(1H, m), 7.98(3H, br),9.57(2H, br s), 12.97(1H, br). 34-14

375.86 340 A (500 MHz, DMSO-d6): 0.26-0.37(2H, m), 0.44-0.55(2H, m),1.11-1.20(1H, m), 2.95(2H, br s), 3.63(2H, br s), 3.84 (2H, d, J=6.9Hz),6.51-6.72(3H, m), 7.26(1H, t, J=8.4Hz), 7.75-8.35(6H, m), 10.39(1H, br),12.89(1H, br). 34-15

465.99 430 A A (500 MHz, DMSO-d6): 0.26-0.37(2H, m), 0.44-0.55(2H, m),1.10-1.22(1H, m), 3.11(2H, br s), 3.74(2H, br s), 3.85 (2H, d, J=6.6Hz),4.18(2H, br s), 6.50- 6.74(3H, m), 7.22-7.32(1H, m), 7.39- 7.48(3H, m),7.52-7.58(2H, m), 7.86 and 8.21(2H, br), 9.21 and 10.35(2H, #br),12.81(1H, br). 34-16

443.51 444 A A (500 MHz, DMSO-d6): 0.25-0.37(2H, m), 0.47-0.57(2H, m),1.22-1.33(1H, m), 3.41-3.54(4H, m), 3.83(2H, d, J= 6.9Hz), 6.39-6.49(2H,m), 6.60-7.00 (3H, m), 7.14(1H, t, J=8.2Hz), 7.23 (1H, br s), 7.43(2H,t, J=7.3Hz), 7.51 (1H, t, J=7.3Hz), 7.80(2H, d, J=7.3 #Hz), 8.62(1H, brs), 14.39(1H, br). 34-17

479.56 480 A C (500 MHz, DMSO-d6): 0.25-0.36(2H, m), 0.47-0.58(2H, m),1.18-1.30(1H, m), 2.96(2H, q, J=6.0Hz), 3.33(2H, q, J=6.0Hz), 3.82(2H,d, J=6.9Hz), 6.40- 6.48(2H, m), 6.60(1H, t, J=6.0Hz), 6.76(2H, s),7.07(1H, s), 7.13(1H, t, J= 8.2Hz), 7.52(2H, t, J=7.3Hz), 7.59 #(1H, t,J=7.3Hz), 7.75(2H, d, J=7.3 Hz), 7.80(1H, t, J=6.0Hz), 14.28(1H, br s).34-18

458.52 459 A A (500 MHz, DMSO-d6): 0.26-0.37(2H, m), 0.50-0.62(2H, m),1.22-1.34(1H, m), 3.35(4H, br s), 3.83(2H, d, J=6.9 Hz), 6.33(1H, br s),6.40-6.48(2H, m), 6.80(2H, br), 6.85-6.92(1H, m), 7.10- 7.25(4H, m),7.37(2H, d, J=7.6Hz), 8.57(1H, s), 14.44(1H, br). 34-19

381.44 382 A A (300 MHz, CDCl3): 0.31-0.41(2H, m), 0.60-0.70(2H, m),1.21-1.38(1H, m), 1.96(3H, s), 3.40-3.50(4H, m), 3.84 (2H, d, J=7.2Hz),5.60(2H, br s), 6.11 (1H, br s), 6.37(1H, d, J=8.3Hz), 6.53 (1H, d,J=8.3Hz), 7.13(1H, t, J=8.3 Hz), 7.35(1H, s), 7.48(1H, br s), 14.44#(1H, br s).

Example 35-1

To a suspension of potassium tert-butoxide (0.809 g, 6.128 mmol) in THF(5 mL) was added a solution of1-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]-phenyl}ethanone (1.00g, 3.064 mmol)(starting compound 1G) in THF (5 mL) followed by carbondisulfide (0.23 mL, 3.83 mmol) and methyl iodide (0.57 mL, 9.19 mmol)successively. The resulting mixture was stirred at room temperature for40 min, and then partitioned between ethyl acetate and water. Theorganic phase was washed with brine, dried over Na₂SO₄, filtered, andconcentrated under reduced pressure. Purification by columnchromatography on silica gel (hexane/ethyl acetate 4:1 to 3:1) gave1-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-3,3-bis(methylsulfanyl)-2-propene-1-oneas a pale yellow oil (0.825 g, yield; 63%).

To a suspension of NaH (60%, 0.056 g, 1.39 mmol) in TBF (3 mL) was addeda solution of t-butyl cyanoacetate (0.197 g, 1.392 mmol) in THF (1 mL)in one portion. After 10 min, a solution of1-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)-oxy]phenyl}-3,3-bis(methylsulfanyl)-2-propene-1-one(0.500 g, 1.16 mmol) in THF (3 mL) was added followed bydibenzo-18-crown-6 (0.013 g, 0.035 mmol). The resulting mixture wasrefluxed for 4 hrs. After cooled to room temperature, the mixture waspartitioned between 0.1 N acetic acid solution and CH₂Cl₂. The organicphase was separated, dried over Na₂SO₄, filtered, and concentrated underreduced pressure. The residual oil was dissolved in 1,2-dichloroethane(1 mL) and acetic acid (1 mL), and ammonium acetate (1.20 g, 15.57 mmol)was added. The mixture was stirred at 120° C. for 0.5 hrs. After cooledto room temperature, the mixture was partitioned between ethyl acetateand saturated aqueous NaHCO₃ solution. The organic phase was washed withsaturated aqueous NaHCO₃ solution and brine, dried over Na₂SO₄,filtered, and concentrated under reduced pressure. Purification bycolumn chromatography on silica gel (hexane/ethyl acetate 4:1 to 3:1)gave tert-butyl2-amino-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-4-(methyl-sulfanyl)nicotinateas an pale yellow oil (0.177 g, yield; 29%).

To a cold (0° C.) solution of tert-butyl2-amino-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-4-(methylsulfanyl)nicotinate(1.72 g, 3.30 mmol) in CH₂Cl₂ (20 mL) was added m-CPBA (69%, 0.82 g,3.30 mmol). The reaction mixture was stirred at 0° C. for 15 min,quenched with saturated aqueous NaHCO₃ solution (20 mL) containingNa₂S₂O₃ (1 g). The organic phase was separated, dried over Na₂SO₄,filtered, and concentrated under reduced pressure. Purification bycolumn chromatography on silica gel (hexane/ethyl acetate=3:2 to 1:1)gave solid, which was triturated with ethyl acetate and diisoproyl etherto give tert-butyl2-amino-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-4-(methylsul-finyl)nicotinateas a white powder (1.112 g, yield; 63%).

A mixture of tert-butyl2-amino-6-{2-(cyclopropylmethoxy)-6-[(4-methoxy-benzyl)oxy]phenyl}-4-(methylsulfinyl)nicotinate(0.118 g, 0.219 mmol) and ethylenediamine (0.29 mL) was stirred at 100°C. for 8 hrs. After cooled to room temperature, the mixture waspartitioned between ethyl acetate and water. The separated organic phasewas washed with brine, dried over Na₂SO₄, filtered, and concentratedunder reduced pressure. The residue was dissolved in CH₂Cl₂ (2 mL), anddi-tert-butyl dicarbonate (0.060 g, 0.275 mmol) was added. The mixturewas stirred at room temperature for 2 hrs and concentrated under reducedpressure. The residue was purified by column chromatography to givetert-butyl2-amino-4-({2-[(tert-butoxycarbonyl)amino]ethyl}amino)-6-{2-(cyclopropylmethoxy)-6-[(4-methoxy-benzyl)oxy]phenyl}nicotinateas a foam (0.121 g, 92%).

To a cold (0° C.) solution of tert-butyl2-amino-4-((2-[(tert-butoxycarbonyl)-amino]ethyl}amino)-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-nicotinate(0.113 g, 0.178 mmol) in THF (5 mL) was added a solution of Vitride®(3.4 M, 1.0 mL, 3.4 mmol). The mixture was stirred at 0° C. for 0.5 hrs,quenched with saturated aqueous potassium sodium tartaric acid solution,and then partitioned between ethyl acetate and water. The organic phasewas washed with brine, dried over Na₂SO₄, filtered, and concentratedunder reduced pressure to give a solid, which was triturated with ethylacetate and diisopropyl ether to give tert-butyl2-{[2-amino-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]-phenyl}-3-(hydroxymethyl)-4-pyridinyl]amino}ethylcarbamateas a pink solid (0.078 g, yield; 78%).

To a cold (0° C.) solution of tert-butyl2-{[2-amino-6-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)oxy]phenyl}-3-(hydroxymethyl)-4-pyridinyl]amino}ethyl-carbamate(0.050 g, 0.089 mmol) and Et₃N (0.074 mL, 0.531 mmol) in THF (10 mL) wasadded triphosgene (0.013 g, 0.044 mmol) in one portion. The mixture wasstirred at 0° C. for 1 hr, and then partitioned between ethyl acetateand water. The organic phase was washed with brine, dried over Na₂SO₄,filtered, and concentrated under reduced pressure. Purification bycolumn chromatography on silica gel (ethyl acetate) gave tert-butyl2-[(7-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)-oxy]phenyl}-2-oxo-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-5-yl)amino]ethylcarbamateas colorless oil (0.036 g, yield; 69%).

To a solution of tert-butyl2-[(7-{2-(cyclopropylmethoxy)-6-[(4-methoxybenzyl)-oxy]phenyl}-2-oxo-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-5-yl)amino]ethyl-carbamate(0.032 g, 0.054 mmol) in 1,4-dioxane (1 mL) was added 4N HCl in1,4-dioxane (1 mL). The mixture was stirred at room temperatureovernight. The resulting precipitate was collected by filtration, washedwith ethyl acetate to give5-[(2-aminoethyl)amino]-7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-2-onehydrochloride as a solid (0.013 g, yield; 59%).

Molecular weight: 406.87; Mass spectrometry: 370 (M+H)⁺; In vitroactivity grade: A; Cellular activity grade: (A549)-A; ¹H-NMR (500 MHz,DMSO-d6): 0.26-0.36 (2H, m), 0.47-0.57 (2H, m), 1.15-1.25 (1H, m),2.96-3.07 (2H, m), 3.50-3.60 (2H, m), 3.85 (2H, d, J=6.9 Hz), 5.45 (2H,br s), 6.57 (1H, d, J=8.2 Hz), 6.56-6.68 (1H, m), 6.96 (1H, br s), 7.24(1H, t, J=8.2 Hz), 7.75 (1H, br), 8.18 (3H, br s), 10.86 (1H, br).

Example 35-2

In a similar manner as that of Example 35-1,5-(2-benzylamino-ethylamino)-6-(2-cyclopropylmethoxy-6-hydroxy-phenyl)-1,4-dihydro-pyrido[2,3-d][1,3]oxazin-2-onehydrochloride was prepared.

Molecular weight: 497.00; Mass spectrometry: 461 (M+H)⁺; In vitroactivity grade: A; (500 MHz, DMSO-d6): 0.26-0.35 (2H, m), 0.45-0.58 (2H,m), 1.10-1.28 (1H, m), 3.16 (2H, br s), 3.67 (2H, br), 3.85 (2H, d,J=6.8 Hz), 4.13-4.25 (2H, m), 5.43 (2H, br s), 6.50-6.65 (2H, m), 7.04(1H, br s), 7.22 (1H, t, J=8.3 Hz), 7.35-7.50 (4H, m), 7.50-7.65 (3H,m), 9.44 (2H, br), 10.74 (1H, br).

What is claimed is:
 1. A pyridine derivative of the formula (I)

wherein —R¹ represents

in which R¹¹ is hydrogen, C₁₋₆ alkyl, halogen, hydroxy, C₁₋₁₂ alkoxy,nitro, amino, C₁₋₆ alkylsulfonylamino, C₁₋₆ alkoxycarbonyl, C₁₋₆alkylamino, di(C₁₋₆ alkyl)amino, C₁₋₆ alkanoylamino, phenyl C₁₋₆alkylamino, phenylsulfonylamino, or —O—(CH₂)_(n)—R¹¹¹, wherein nrepresents an integer selected from 0 to 6, and R¹¹¹ is C₂₋₆ alkenyl,benzoyl, diphenylmethyl, di(C₁₋₆ alkyl)amino, C₁₋₆ alkanoyl, C₁₋₆alkoxycarbonyl, or a 3 to 10 membered saturated or unsaturated ringhaving 0 to 3 heteroatoms selected from the group consisting of S, O andN as heteroatoms and is optionally substituted by C₁₋₆ alkyl, mono or dihalogen, halogen substituted C₁₋₆ alkyl, nitro, cyano, C₁₋₆alkoxycarbonyl, phenyl, hydroxy, amino, C₁₋₆ alkylamino, di(C₁₋₆alkyl)amino, C₁₋₆ alkanoylamino, C₁₋₆ alkoxy, or carbamoyl; R²represents hydrogen or halogen; R³ represents hydrogen or1,2,3,6-Tetrahydro-pyridine, —CR³¹R³²R³³,  wherein R³¹ is hydrogen orC₁₋₆ alkyl, R³² is hydrogen, α-aminobenzyl, C₁₋₆ alkyl optionallysubstituted by one or two substituents selected from the groupconsisting of hydroxy, amino, amino substituted phenyl, phenyl, halogensubstituted phenyl, and C₁₋₆ alkoxysubstituted phenyl, or a 5 to 8membered saturated ring having 0 to 3 atoms selected from the groupconsisting of S, O and N as heteroatoms and optionally substituted byC₁₋₆ alkyl, and R³³ is hydrogen, amino, C₁₋₆ alkoxycarbonylamino, C₂₋₆alkenyloxycarbonylamino, piperidino-C₁₋₆ alkylcarbonylamino,piperidinyl-C₁₋₆ alkylcarbonylamino, or R³² and R³³ may form, togetherwith the adjacent carbon atom, a 5 to 8 membered saturated ring having 0to 3 heteroatoms selected from the group consisting of N, O and S asheteroatoms, which ring is optionally substituted by phenyl-C₁₋₆ alkyl,C₁₋₆ alkoxy substituted phenyl-C₁₋₆ alkyl, C₁₋₆ alkyl, amino, cyano,hydroxy, carbamoyl, carboxy, C₁₋₆ alkylamino, C₁₋₆ alkoxycarbonyl,di(C₁₋₆ alkyl)amino, benzylamino, C₁₋₆ alkylsulfonyl, piperidino C₁₋₆alkyl carbonyl, or optionally fused by benzene; or —NR³⁴R³⁵,  whereinR³⁴ is hydrogen or C₁₋₆ alkyl and R³⁵ is hydrogen, a 5 to 8 memberedsaturated ring having 0 to 3 heteroatoms selected from the groupconsisting of N, O and S as heteroatoms, or —(CH₂)_(m)—NR³⁵¹R³⁵² (mrepresents any of integers from 1 to 6) wherein R³⁵¹ representshydrogen, C₁₋₆ alkyl, R³⁵² represents hydrogen, C₁₋₆ alkyl, C₁₋₆alkanoyl, C₁₋₆ alkylsubstituted phenyl, benzoyl, C₁₋₆ alkanoyl,phenylaminocarbonyl, phenylsulfonyl, or R³⁴ and R³⁵ may form, togetherwith the adjacent N atom, a 5 to 8 membered saturated heterocyclic ring,and said ring may optionally contain NH, S or O atom other than theadjacent N atom and optionally substituted by carbamoyl, amino, or C₁₋₆alkyl; R⁴ and R₅ form, —R⁴⁰—CO—NH—, —R⁴⁰—SO₂—NH—, —R⁴⁰—C(═S)—NH——R⁴⁰—CH₂—NH—,  wherein said —R⁴⁰— represents —CHR⁴⁰¹—O—, —CH₂—NR⁴⁰¹—,—CO—NR⁴⁰¹—, —CH₂—CHR⁴⁰¹—, —CH═CR⁴⁰¹—, (in which R⁴⁰¹ is C₁₋₆ alkanoyl,C₁₋₆ alkyl, phenyl, C₁₋₆ alkylsulfonyl, C₃₋₈ cycloalkylaminocarbonyl,hydrogen, halogen, nitro, amino, cyano, benzoylamino, phenylsulfonyl,carbamoyl, hydroxycarbonyl, C₁₋₆ alkoxycarbonyl, C₁₋₁₂alkylaminocarbonyl, halogen substituted C₁₋₆ alkylaminocarbonyl, C₁₋₆alkanoylamino, C₁₋₆ alkylamino, di(C₁₋₆ alkyl)aminocarbonyl, di(C₁₋₆alkyl)aminoC₁₋₆ alkylaminocarbonyl, hydroindenylaminocarbonyl,diphenylmethylaminocarbonyl, pyrrolidinocarbonyl, C₁₋₆ alkoxy C₁₋₆ alkylamino carbonyl, morpholinocarbonyl, piperazinocarbonyl,phenylC₁₋₆alkylaminocarbonyl, hydroxycarbonylC₁₋₆alkylaminocarbonyl,C₃₋₈ cycloalkylaminocarbonyl, C₃₋₈ cycloalkylC₁₋₆alkylaminocarbonyl,hydroxyC₁₋₆alkylaminocarbonyl, carboxyethylaminocarbonyl,C₁₋₆alkylsulfonylaminocarbonyl) —CR⁴¹═N—NH— (R⁴¹ is hydrogen, amino, orC₁₋₆alkanoylamino), or —CR⁴²═N—C═N— (R⁴² is hydrogen or amino); andsalts thereof.
 2. The compound or a salt thereof as claimed in claim 1,wherein: —R¹ represents

in which R¹¹ is hydrogen, C₁₋₆ alkyl, halogen, hydroxy, C₁₋₁₂ alkoxy,amino, C₁₋₆ alkanoylamino, phenyl C₁₋₆ alkylamino, phenylsulfonylamino,or —O—(CH₂)_(n)—R¹¹¹, wherein n represents an integer selected from 1 to6, and R¹¹¹, is C₂₋₆ alkenyl, benzoyl, diphenylmethyl, di(C₁₋₁₆alkyl)amino, C₁₋₆ alkanoyl, C₁₋₆ alkoxycarbonyl, or a 3 to 10 memberedsaturated or unsaturated ring having 0 to 3 heteroatoms selected fromthe group consisting of S, O and N as heteroatoms and is optionallysubstituted by C₁₋₆ alkyl, mono or di halogen, halogen substituted C₁₋₆alkyl, nitro, cyano, C₁₋₆ alkoxycarbonyl, phenyl; R² representshydrogen; R³ represents hydrogen, 1,2,3,6-tetrahydro-pyridine—CR³¹R³²R³³,  wherein R³¹ is hydrogen or C₁₋₆ alkyl, R³² is hydrogen,α-aminobenzyl, C₁₋₆ alkyl optionally substituted by one or twosubstituents selected from the group consisting of hydroxy, amino, aminosubstituted phenyl, phenyl, halogen substituted phenyl, and C₁₋₆alkoxysubstituted phenyl, or a 5 to 8 membered saturated ring having 0to 3 atoms selected from the group consisting of S, O and N asheteroatoms and optionally substituted by C₁₋₆ alkyl, and R³³ ishydrogen, amino, C₁₋₆alkoxycarbonylamino, C₂₋₆ alkenyloxycarbonylamino,piperidino-C₁₋₆ alkylcarbonylamino, or R³² and R³³ may form, togetherwith the adjacent carbon atom, a 5 to 8 membered saturated ring having 0to 3 heteroatoms selected from the group consisting of N, O and S asheteroatoms, which ring is optionally substituted by phenyl-C₁₋₆ alkyl,C₁₋₆ alkoxy substituted phenyl-C₁₋₆ alkyl, C₁₋₆ alkyl, amino, carboxy,C₁₋₆ alkylamino, C₁₋₆ alkoxycarbonyl, di(C₁₋₆ alkyl)amino, benzylamino,C₁₋₆ alkylsulfonyl, piperidino C₁₋₆ alkyl carbonyl, or optionally fusedby benzene; or —NR³⁴R³⁵,  wherein R³⁴ is hydrogen and R³⁵ is hydrogen, a5 to 8 membered saturated ring having 0 to 3 heteroatoms selected fromthe group consisting of N, O and S as heteroatoms, or—(CH₂)_(m)—NR³⁵¹R³⁵² (m represents any of integers from 1 to 6) whereinR³⁵¹ represents hydrogen, C₁₋₆ alkyl, R³⁵² represents hydrogen, C₁₋₆alkyl, C₁₋₆ alkanoyl, C₁₋₆ alkylsubstituted phenyl, benzoyl, C₁₋₆alkanoyl, phenylaminocarbonyl, phenylsulfonyl, or R³⁴ and R³⁵ may form,together with the adjacent N atom, a 5 to 8 membered saturatedheterocyclic ring, and said ring may optionally contain NH, S or O atomother than the adjacent N atom and optionally substituted by carbamoyl,amino, or C₁₋₆ alkyl; R⁴ and R⁵ form, —R⁴⁰—CO—NH—, —R⁴⁰—SO₂—NH—,—R⁴⁰—C(═S)—NH—, —R⁴⁰—CH₂—NH—,  wherein said —R⁴⁰— represents —CHR⁴⁰¹—O—,—CH₂—NR⁴⁰¹—, —CO—NR⁴⁰¹—, (in which R⁴⁰¹ is hydrogen, C₁₋₆ alkanoyl, C₁₋₆alkoxycarbonyl, C₁₋₆ alkyl, phenyl, C₁₋₆ alkylsulfonyl, C₃₋₈cycloalkylaminocarbonyl, C₁₋₆ alkylaminocarbonyl, carbamoyl, di(C₁₋₆alkyl)aminocarbonyl), —CH₂—CHR⁴⁰²—, —CH═CR⁴⁰²—, (in which R⁴⁰² ishydrogen, halogen, nitro, amino, cyano, benzoylamino, phenylsulfonyl,carbamoyl, hydroxycarbonyl, C₁₋₆ alkoxycarbonyl, C₁₋₁₂alkylaminocarbonyl, halogen substituted C₁₋₆ alkylaminocarbonyl, C₁₋₆alkanoylamino, C₁₋₆ alkylamino, di(C₁₋₆ alkyl)aminocarbonyl, di(C₁₋₆alkyl)aminoC₁₋₆ alkylaminocarbonyl, hydroindenylaminocarbonyl,diphenylmethylaminocarbonyl, pyrrolidinocarbonyl, C₁₋₆ alkoxy C₁₋₆ alkylamino carbonyl, morpholinocarbonyl, piperazinocarbonyl, phenylC₁₋₆alkylaminocarbonyl, C₃₋₈cycloalkylaminocarbonyl,hydroxycarbonylC₁₋₆alkylaminocarbonyl, C₃₋₈cycloalkylC₁₋₆alkylaminocarbonyl, hydroxyC₁₋₆alkylaminocarbonyl,carboxyethylaminocarbonyl, methylsulfonylaminocarbonyl,) —CR⁴¹═N—NH—(R⁴¹ is hydroxy, amino, C₁₋₆ alkanoylamino) or —CR⁴²═N—C═N— (R⁴² isamino).
 3. The compound or a salt thereof as claimed in claim 1,wherein: —R¹ represents

in which R¹¹ is hydrogen, C₁₋₁₂ alkoxy, or —O—(CH₂)_(n)—R¹¹¹, wherein nrepresents an integer selected from 1 to 6, and R¹¹¹ is phenyl, C₃₋₈cycloalkyl; R² represents hydrogen; R³ represents1,2,3,6-tetrahydro-pyridine, —CR³¹R³²R³³,  wherein R³¹ is hydrogen, andR³² and R³³ form, together with the adjacent carbon atom, a 5 to 8membered saturated ring interrupted by NH, which ring is optionallysubstituted by phenyl-C₁₋₆ alkyl, C₁₋₆ alkoxy substituted phenyl-C₁₋₆alkyl, C₁₋₆alkyl, amino, carboxy, C₁₋₆ alkylamino, C₁₋₆ alkoxycarbonyl,di(C₁₋₆ alkyl)amino, benzylamino, C₁₋₆ alkylsulfonyl, piperidino C₁₋₆alkyl carbonyl, or optionally fused by benzene; or —NR³⁴R³⁵,  whereinR³⁴ is hydrogen and R³⁵ is —(CH₂)_(m)—NR³⁵¹R³⁵² (m represents any ofintegers from 1 to 6) wherein R³⁵¹ represents hydrogen, C₁₋₆ alkyl, R³⁵²represents hydrogen, C₁₋₆ alkyl, C₁₋₆ alkanoyl, C₁₋₆ alkylsubstitutedphenyl, benzoyl, C₁₋₆ alkanoyl, phenylaminocarbonyl, phenylsulfonyl; andR⁴ and R⁵ form, —R⁴⁰—CO—NH—, —R⁴⁰—SO₂—NH—, —R⁴⁰—C(═S)—NH—, —R⁴⁰—CH₂—NH—, wherein said —R⁴⁰— represents —CHR⁴⁰¹—O—, —CH₂—NR⁴⁰¹—, —CO—NR⁴⁰¹—, (inwhich R⁴⁰¹ is hydrogen, C₁₋₆ alkanoyl, C₁₋₆ alkoxycarbonyl, C₁₋₆ alkyl,phenyl, C₁₋₆ alkylsulfonyl, C₃₋₈ cycloalkylaminocarbonyl, C₁₋₆alkylaminocarbonyl, carbamoyl, di(C₁₋₆ alkyl)aminocarbonyl),—CH₂—CHR⁴⁰²—, —CH═CR⁴⁰²—, (in which R⁴⁰² is hydrogen, halogen, nitro,amino, cyano, benzoylamino, phenylsulfonyl, carbamoyl, hydroxycarbonyl,C₁₋₆ alkoxycarbonyl, C₁₋₁₂ alkylaminocarbonyl, halogen substituted C₁₋₆alkylaminocarbonyl, C₁₋₆ alkanoylamino, C₁₋₆ alkylamino, di(C₁₋₆alkyl)aminocarbonyl, di(C₁₋₆ alkyl)aminoC₁₋₆ alkylaminocarbonyl,hydroindenylaminocarbonyl, diphenylmethylaminocarbonyl,pyrrolidinocarbonyl, C₁₋₆ alkoxy C₁₋₆ alkyl amino carbonyl,morpholinocarbonyl, piperazinocarbonyl, phenylC₁₋₆alkylaminocarbonyl,C₃₋₈ cycloalkylaminocarbonyl, hydroxycarbonylC₁₋₆ alkylaminocarbonyl,C₃₋₈ cycloalkylC₁₋₆alkylaminocarbonyl, hydroxyC₁₋₆ alkylaminocarbonyl,carboxyethylaminocarbonyl, methylsulfonylaminocarbonyl,) —CR⁴¹═N—NH—(R⁴¹is hydroxy, amino, C₁₋₆alkanoylamino) or —CR⁴²═N—C═N— (R⁴² is amino). 4.The compound or a salt thereof as claimed in claim 1, wherein —R¹represents

in which R¹¹ is hydrogen, C₁₋₆ alkoxy, or —O—CH₂—R¹¹¹, wherein R¹¹¹ isphenyl, C₃₋₄ cycloalkyl; R² represents hydrogen; R³ represents1,2,3,6-tetrahydro-pyridine, —CR³¹R³²R³³,  wherein R³¹ is hydrogen, andR³² and R³³ form, together with the adjacent carbon atom, piperidine orpyrrolidine, which piperidine or pyrrolidine is optionally substitutedby C₁₋₆ alkyl; or —NR³⁴R³⁵,  wherein R³⁴ is hydrogen and R³⁵ is—(CH₂)_(m)—NR³⁵¹R³⁵² (m represents any of integers from 1 to 6) whereinR³⁵¹ represents hydrogen, C₁₋₆ alkyl, R³⁵² represents hydrogen, C₁₋₆alkyl, C₁₋₆ alkanoyl, C₁₋₆ alkylsubstituted phenyl, benzoyl, C₁₋₆alkanoyl, phenylaminocarbonyl, phenylsulfonyl; and R⁴ and R⁵ form,—R⁴⁰—CO—NH— or —R⁴⁰—CH₂—NH—,  wherein said —R⁴⁰— represents —CHR⁴⁰¹—O—,—CH₂—NR⁴⁰¹—, —CO—NR⁴⁰¹—, (in which R⁴⁰¹ is hydrogen, C₁₋₆ alkanoyl, C₁₋₆alkoxycarbonyl, C₁₋₆ alkyl, phenyl, C₁₋₆ alkylsulfonyl, C₃₋₈cycloalkylaminocarbonyl, C₁₋₆ alkylaminocarbonyl, carbamoyl, di(C₁₋₆alkyl)aminocarbonyl), —CH₂—CHR⁴⁰²— or —CH═CR⁴⁰²—, (in which R⁴⁰² ishydrogen, halogen, nitro, amino, cyano, benzoylamino, phenylsulfonyl,carbamoyl, hydroxycarbonyl, C₁₋₆ alkoxycarbonyl, C₁₋₁₂alkylaminocarbonyl, halogen substituted C₁₋₆ alkylaminocarbonyl, C₁₋₆alkanoylamino, C₁₋₆ alkylamino, di(C₁₋₆ alkyl)aminocarbonyl, di(C₁₋₆alkyl)aminoC₁₋₆ alkylaminocarbonyl, hydroindenylaminocarbonyl,diphenylmethylaminocarbonyl, pyrrolidinocarbonyl, C₁₋₆ alkoxy C₁₋₆ alkylamino carbonyl, morpholinocarbonyl, piperazinocarbonyl,phenylC₁₋₆alkylaminocarbonyl, C₃₋₈ cycloalkylaminocarbonyl,hydroxycarbonylC₁₋₆alkylaminocarbonyl, C₃₋₈cycloalkylC₁₋₆alkylaminocarbonyl, hydroxyC₁₋₆alkylaminocarbonyl,carboxyethylaminocarbonyl, methylsulfonylaminocarbonyl).
 5. The compoundas claimed in claim 1 selected from the group consisting of thefollowing compounds:7-(2-hydroxyphenyl)-5-(3-piperidinyl)-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-2-one;7-[2-(benzyloxy)-6-hydroxyphenyl]-5-(3-piperidinyl)-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-2-one;7-(2-hydroxyphenyl)-5-(3-piperidinyl)-3,4-dihydro-1,8-naphthyridin-2(1H)-one;7-(2-hydroxy-6-propoxyphenyl)-5-(3-piperidinyl)-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-2-one;ethyl7-(2-hydroxy-6-propoxyphenyl)-2-oxo-5-(3-piperidinyl)-1,2,3,4-tetrahydro-1,8-naphthyridine-3-carboxylate;7-(2-hydroxy-6-propoxyphenyl)-5-(3-piperidinyl)-3,4-dihydro-1,8-naphthyridin-2(1H)-one;7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-5-(3-piperidinyl)-3,4-dihydro-1,8-naphthyridin-2(1H)-one;ethyl7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-5-(3-piperidinyl)-1,2,3,4-tetrahydro-1,8-naphthyridine-3-caboxylate;7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-5-(4-piperidinyl)-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-2-one;7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-5-(4-piperidinyl)-3,4-dihydro-1,8-naphthyridin-2(1H)-one;7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-3-fluoro-5-(3-piperidinyl)-1,8-naphthyridin-2(1H)-one;7-(2-hydroxy-6-propoxyphenyl)-5-(4-piperidinyl)-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-2-one;7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-5-(3-piperidinyl)-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-2-one;3-(cyclopropylmethoxy)-2-[5-(3-piperidinyl)-1,4-dihydro-2H-pyrido[2,3-d][1,3]oxazin-7-yl]phenol;7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-5-(3-piperidinyl)-1,8-naphthyridin-2(1H)-one;7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-5-(3-piperidinyl)-1,2-dihydro-1,8-naphthyridine-3-carboxamide;3-acetyl-7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-5-(3-piperidinyl)-3,4-dihydropyrido[2,3-d]pyrimidin-2(1H)-one;7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-5-(3-piperidinyl)-3,4-dihydropyrido[2,3-d]pyrimidin-2(1H)-one;7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-5-(3-piperidinyl)-1,2,3,4-tetrahydro-1,8-naphthyridine-3-carboxamide;7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-N-isopropyl-2-oxo-5-(3-piperidinyl)-1,2-dihydro-1,8-naphthyridine-3-carboxamide;ethyl7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-5-(3-piperidinyl)-1,4-dihydropyrido[2,3-d]pyrimidine-3(2H)-carboxylate;7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-N,N-dimethyl-2-oxo-5-(3-piperidinyl)-1,4-dihydropyrido[2,3-d]pyrimidine-3(2H)-carboxamide;7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-5-(3-piperidinyl)-1,4-dihydropyrido[2,3-d]pyrimidine-3(2H)-carboxamide;isopropyl7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-5-(3-piperidinyl)-1,4-dihydropyrido[2,3-d]pyrimidine-3(2H)-carboxylate;isobutyl7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-5-(3-piperidinyl)-1,4-dihydropyrido[2,3-d]pyrimidine-3(2H)-carboxylate;neopentyl7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-2-oxo-5-(3-piperidinyl)-1,4-dihydropyrido[2,3-d]pyrimidine-3(2H)-carboxylate;and7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-N-ethyl-2-oxo-5-(3-piperidinyl)-1,4-dihydropyrido[2,3-d]pyrimidine-3(2H)-carboxamide;and salts thereof.
 6. A medical composition comprising the compound or asalt thereof as claimed in claim 1 together with one or morepharmaceutically acceptable excipients.
 7. A method of inhibiting IκBkinase β in a mammal, comprising administering a compound or a saltthereof as claimed in claim
 1. 8. A method of treating an inflammatorydisease in a mammal, comprising administering a compound or a saltthereof as claimed in claim
 1. 9. The method as claimed in claim 8,wherein said inflammatory disease is selected from the group consistingof asthma, allergic rhinitis, atopic dermatitis, hives, conjunctivitis,vernal catarrh, chronic arthrorheumatism, systemic lupus erythematosus,psoriasis, diabrotic colitis, systemic inflammatory response syndrome(SIRS), sepsis, polymyositis, dermatomyositis (DM), Polyaritis nodoa(PN), mixed connective tissue disease (MCTD), Sjoegren's syndrome andgout.
 10. A method of promoting immunosuppression in a mammal,comprising administering a compound or a salt thereof as claimed inclaim
 1. 11. A method of treating ischemia in a mammal, comprisingadministering a compound or a salt thereof as claimed in claim
 1. 12. Amethod of treating tumors in a mammal, comprising administering acompound or a salt thereof as claimed in claim 1.